1. science
2. medicine
3. immunology

Department of Microbiology,
Tumor and Cell Biology
Scientific Report 2007–2009
2
Department of Microbiology, Tumor and Cell Biology – MTC
Contents
Looking Beyond Tomorrow – Moving into the Future.............................4
MTC 15 Years – Where We Came From...............................................6
Newcomers
A Long and Winding Road to “Chromosomics”...............................8
Bringing a Touch of Chemistry to MTC..........................................10
Tracking Down Fresh Footprints of the Myc Tumor Protein..........11
The Secret Garden of Rho GTPases............................................12
Awards, Major Grants and Appointments............................................14
Scientists in Focus
From the Clinics to Basic Science.................................................18
Towards an Adjunct Professorship in Parasitology ......................19
Hans G Boman in Memoriam .............................................................20
High Impact Publications 2007–2009.................................................23
Immunology Research........................................................................28
Companies Founded by MTC Scientists
Intervacc AB.................................................................................84
DILAFOR AB................................................................................84
Future Faculty.....................................................................................85
Equality and Diversity Group..............................................................85
Undergraduate Education...................................................................86
Training for the Future – PhD Studies.................................................87
Summer Research Schools................................................................88
Life as a PhD Student.........................................................................89
Doctoral Theses 2007–2009...............................................................90
Core Facilities
MTC FACS Facility.......................................................................94
KI Visualization Facility (KIVIF).....................................................95
MTC Research Facility ................................................................96
Infection Biology Research.................................................................38
HIV-laboratory .............................................................................97
Cell and Tumor Biology Research.......................................................56
Interactions with the Outside World.....................................................98
What Happens Down in the Cellar? Lennart Nilsson at MTC..............68
Georg and Eva Klein Foundation........................................................99
Foreign Adjunct Professors
MTC Organization............................................................................100
p53 Forever – Focus on this Fascinating Protein..........................70
MTC – Not Only a Scientific Experience!......................................71
Innovating Vaccines by Translational Entrepreneurship...............72
Our valuable Collaboration with the Karolinska Institutet..............73
MTC International
Economy..........................................................................................101
Contributors and List of Financiers...................................................102
Location of MTC...............................................................................103
Evaluation of HIV Vaccines in Tanzania........................................74
Scientific Alliances in Africa
.........................................................75
The Singapore Partnership for the Future....................................76
International networks for high-quality graduate education..........77
Scientific Networks
KICancer......................................................................................78
Editor-in-Chief: Marie Arsenian Henriksson
Scientific Editors: Mikael Jondal and Ute Römling
Bibliometric Analysis: Catharina Rhen
Text Editor: Anna Lögdberg
Photo: Fredrik Persson and John Sennett
KI-Infection...................................................................................79
Artistic Contributions: Ewert Linder
KI Inflammation and Immunology.................................................79
Layout: Björn Lundquist, Malmö
Print: Edita Västra Aros
Emerging Collaborations
The Royal Institute of Technology (KTH)......................................80
Center for Infectious Medicine (CIM)............................................81
Translational Research.......................................................................82
Cover photo by Lennart Nilsson/Scanpix:
Malaria parasites have multiplied inside two red blood
cells. One has burst open.
Department of Microbiology, Tumor and Cell Biology – MTC
3
Head of Department
Looking Beyond Tomorrow
– Moving into the Future
It all starts with an open mind, a new
idea. A mixture of creativity and originality in
combination with intellectual courage, hard
and committed work as well as the intuition
of a single person. Needless to say, the individual scientist is the driving force and the key to
successful medical research. Equally important is the critical mass of fellow scientists in
related or different areas while the infrastructure and state-of-the-art equipment are the
essential pre-requisites enabling ideas to be
tested experimentally. Karolinska Institutet
(KI) has a strong tradition of cross-fertilization between disciplines. These interactions
permeate all research at the Department
of Microbiology, Tumor and Cell Biology
(MTC), which is characterized by collaborations between preclinical and clinical scientists in dynamic national and international
interactions.
ment policy with a transparent procedure to
guarantee the next generation of scientists.
During the last three years MTC has recruited
several new professors, senior lecturers, adjunct and foreign adjunct professors as well as
scientists with external senior research positions. It is particularly rewarding that several
researchers at early stages in their careers have
joined the department. As science is built on
individuals we will pursue recruitments at all
levels with a focus on young and promising
scientists.
We will deepen the interaction with the
clinic in parallel with building of the new
university hospital, NKS (Nya Karolinska
Solna). A significant part of the collaboration
with the clinic occurs with the unit of Clinical Microbiology, which is part of MTC but
located at the Karolinska University Hospital
Solna. This natural connection to the hospital has been very fruitful and is now being
Our focus is the fight against infections and
intensified. At present several clinical trials
cancer. The merge of immunology, infection
are being performed in
biology and cell and tuthe oncology field. These
mor biology provides us
“We have students and
and other contacts will
with a unique platform
researchers from all over the
be developed further to
where fruitful interacglobe who contribute to our
allow the translation of
tions and creative ideas
intellectual platform built
basic research to mediarise and flourish. As
on creativity, originality,
cal care as well as into
illustrated on page 26,
diversity and equlity.”
commercial products
these interactions are
and business. Scientists
also revealed by biblioat MTC are founders
metric analysis. It is our
and board members of several start-up comfuture mission to maintain MTC as a frontpanies, including Aprea, Dilafor, Imed, Index
line department at the international level in
and Intervacc, located within the Karolinska
science and education. To achieve this, we will
Institutet Science Park.
continue to carry out a pro-active recruit-
4
Department of Microbiology, Tumor and Cell Biology – MTC
The scientific infrastructure is of fundamental
importance when facing international competition. We have made strategic investments in
our FACS facility, atomic force microscopy and
live cell imaging equipment. These long-term
commitments in infrastructure will continue.
Moreover, maintaining a top-notch administration and technical support are necessary
pre-requisites to meet future needs. It is also
rewarding that the new Science for Life Laboratory (SciLifeLab), a national resource center
dedicated to large scale research in molecular
biosciences and medicine, is situated close to
MTC. Our projects will be greatly facilitated
through collaborations and discussions with
the SciLifeLab scientists and will benefit by
being integrated into their large scale stateof-the-art technology platforms. During the
recent years we have increased our interactions and collaborations with the Royal Institute of Technology (KTH) both with respect
to research collaborations as well as technical
developments. These strategic contacts will be
even more important in the future.
International collaborations already established by individual researchers, as well as
interactions formally established with Southern China, Vietnam, Uganda, and Singapore,
will be further developed. Several MTC scientists are involved in the unique, international
research education collaboration between
KI and Makerere University in Kampala,
Uganda. The collaboration with Chinese
cancer research started during the ’90s and
has generated many contacts and projects of
common interest. The collaboration started at
the Sun Yat Sen Cancer Center in Guangzhou
(Canton), but has been expanded to several
regions in China. MTC has also a fruitful interaction with Hanoi Medical University in Vietnam through the research education program
“Common Diseases in Vietnam”. Several PhD
students and post-docs from Vietnam have
been working at MTC during the recent years.
A*STAR is our foremost partner in Singapore,
through which education of both Swedish and
Singaporean students are financed.
All of these collaborations have generated
novel knowledge and several joint publications throughout the years.
Training of the next generation of scientists
is one of the most important missions of any
university. Subsequently, research education
both for PhD students and post-docs is the
core of our activities and builds on learning
the trade in the individual research project,
like in the former guilds. It is of fundamental
importance that we keep this tradition alive,
while ensuring a high quality education.
The intensive seminar series that take place
everyday at MTC play a central role in our
scientific life. They focus on our three areas
of research; immunology, infection ­biology
and tumor cell biology and function as a
platform for communication and discussion
of new scientific developments. In combination with the many international conferences
that are being organized by MTC scientists,
the ­seminars constitute an ideal educational
forum and for spreading knowledge.
It is our challenge to maintain MTC as a
prominent institution where scientists open up
new fields of investigation and make discoveries of fundamental importance. Key words
are multidisciplinary, bridging, translational,
national and international collaboration. The
power and greatest asset of MTC is each and
every individual person working at the department. We have students and researchers from
Africa, Asia, Australia, Europe and the Americas who contribute to the intellectual scientific
platform built on creativity, originality, diversity and equality. It is a joy to foster this spirit to
meet future challenges.
Marie Arsenian Henriksson
Department of Microbiology, Tumor and Cell Biology – MTC
5
Inauguration of
MTC by the Minis
ter of Education
and KI Preside
Per Unckel
nt Bengt Samu
elsson (right).
3.
Group Leaders of MTC in 199
MTC 15 Years – Where We Came From
We celebrated our 15th BIRTHDAY in May 2008 with an international top symposium and a warm and cheerful party at the Medical Union (MF)
venue. MTC was born on the first of July 1993 as a result of the launching of the “KI 93” – a major reform at the Karolinska Institutet. More than
100 departments were fused over night to become 30 “storinstitutioner” – BIG departments.
For our department this was unusually
timely. The construction of our new buildings
was already on its way at the site where they
stand today. The decision to create a totally
new department was made after decades of
discussion about expanding the facilities of
the Tumor Biology Department successfully
lead by George and Eva Klein, the founders
of Tumor Biology at KI and vague plans for a
new KI Microbiology Center.
In 1989 this “long-bench” process transformed into accelerated planning partly due
to a new Swedish government with an interest in reforming the university system. In
particular it was due to a strong push from
inside KI from some colleagues, in particular
the KI president, professor Bengt Samuelsson, Hans Wigzell, professor of immunology,
who later became the president of KI, and
the dean, Erling Norrby, professor of virology. Until the last minute it was uncertain
whether this new center should be located on
the KI campus area in Solna or on the KI area
in Huddinge.
The construction of new buildings was
initially independent of the KI 93 reorganization, but in the end these two events
converged. Integrated into the global reor-
6
Department of Microbiology, Tumor and Cell Biology – MTC
ganization of KI, the fusion of five departments, the Departments of Bacteriology,
Infectious Disease Protection, Immunology,
Tumor Biology and Virology in a new building around the corner went smooth, although
these departments were previously spread all
over the KI campus and the Department of
Virology was even located outside the campus
at the former State Bacteriological Laboratory
(SBL) in Huvudsta.
Organization of MTC
Before moving into the new buildings we
joined forces in these five departments and
formed a dozen working groups, made up
by appropriate staff members, to plan for
everything from a joint economy, to heavy
equipment, clean water and common facilities as well as joint shops, like the “Boutique
Plastique” where researchers can buy plastic
material 24 hours a day. We could then move
into our new houses in February 1994 with
a new organization in place. The overriding concept was “The Research Group in the
Middle”, i.e. an effort to plan everything from
laboratories and offices to economy in order
to support the everyday work of the research
groups.
After all, the creativity and production of
the research groups was the basis for generating a solid economy – pretty obvious today
but not in those days when university departments were organized as public authorities
based on state funding. As a result, MTC
developed a highly research oriented environment. The total budget in 1993–94 was
around 50 million Swedish crowns and has
more than trippled over 15 years.
Thirty research groups were identified
within the old departments. To qualify as
a group leader he/she must have a tenured
or 4–6 year research council position. As
a result several young scientists including women were identified as leaders at the
same level as the most established professors.
They all joined in the MTC faculty group
(“forskarkollegiet” in Swedish) which started
off as a consensus group for forming continued strategies.
Interdisciplinary build up
The diversity and flow of research at MTC
is also reflected in its buildings. During the
whole ’90s the MTC buildings were continuously improved. After moving into the new A,
B, C and D-buildings, part of the old bacte-
The builders of MTC Alex Feldötö and Greger Blomquist.
er.
MTC millenium calend
riology houses were replaced by the new
G-wing in 1996. In the last phase, the teaching
building (E) and the bacteriology building (F)
were renovated, ready for use in 1998.
During these events, research groups had
to move around a lot, some groups as much
as three times! In spite of this inconvenience, this continuous change also created
new constellations of research groups with
new neighbours melting together our 4–5
disciplines. The interdisciplinary build up by
MTC became its hallmark that has generated
a lot of novel collaborations and new projects,
although one could always have wished for
more efforts in this direction.
Due to the large number of groups which
were very successful in being awarded external
research grants, the economy of the department became quite stable. Consequently, there
He did everyth
ing himself – th
e first head of
MTC, professo
r Ingemar Ernb
erg, appearing as the depa
rtmental Lucia
.
was money to buy state-of-the-art equipment
which was the basis to form new core facilities
within the department. By the millennium
shift we had a positive balance and funding for
new investments. This development did not
occur without some pain, however, as during
the ’90s the organization of MTC had to be
dramatically restructured. The key concept
was to invest state funds in the research groups
that in turn could generate more money
through external grants. This process required
the dramatic reduction of supportive personnel such as technicians (BMAs), administrators and engineers. Altogether 35 persons had
to leave MTC during 1995-97. With reforming
the KI economy and introducing a new rental
system MTC again went through economic
harsh times shortly after the beginning of
the new millennium. However, with a strong
management the economy has improved and
MTC is thriving again.
Added value of neighbourship
One important development for MTC was the
move of SBL in Huvudsta to the KI Campus,
next door to MTC. SBL was renamed the
Center for Infectious Disease Control (SMI)
in 1997. Along with this fusion came a donation from the Swedish government for six
research chairs in clinical immunology, virology, bacteriology, parasitology, vaccinology
and epidemiology. Most of these professors
became affiliated with MTC through their
research and PhD education.
Over the years significant efforts were
made to optimize the added value of this
neighbourship. In addition, later The European Center for Disease Control (ECDC) was
placed close to MTC and SMI; all three insti-
tutions together form an impressive cluster of
knowledge in the area of microbiology.
A new generation of scientists at MTC
By the end of the ’90s there had been a major
turnover of research groups. Rolf Kiessling,
Klas Wiman, Eva-Maria Fenyö, Lars Holmgren, Hans-Gustav Ljunggren and Joakim
Dillner were some who left for chairs in
other departments and universities. A whole
generation of modern cell biologists and
microbiologists joined MTC. Newcomers
were Staffan Normark, Yihai Cao and Mikael
Rhen, followed by Ute Römling, Sven Pettersson and Markus Maeurer in the early 2000s
and recently by Lars-Gunnar Larsson, Pontus
Aspenström, Rolf Ohlsson and Sonia Laín.
Thus the turnover in the faculty has been
vivid and healthy.
It is also worth to recognize the balancing
input by our professor emeriti like Carl-Ivar
Brändén (1998 –2003) and Hans G Boman
(1997–2007). Recently professor David Lane
joined the team as a foreign adjunct professor
from the UK and Singapore.
By 2000, professor Ingemar Ernberg
(1993–2000) handed over the department
chair to professor Klas Kärre (2000–2002)
followed by professor Mats Wahlgren (2003–
2006) and associate professor Marie Arsenian
Henriksson (2006–). MTC moved from creation to consolidation. The Microbiology and
Tumor Biology Center (MTC) became The
Department of Microbiology, Tumor and
Cell Biology (MTC) well poised to supply a
creative research environment for young and
senior scientist to make important contributions to human health in the future!
Ingemar Ernberg
Department of Microbiology, Tumor and Cell Biology – MTC
7
Newcomers
Rolf Ohlsson
A Long and Winding Road to “Chromosomics”
as a scientist you
have to show passion for your work. However,
this passion must be something extraordinary to be able to buffer against the disappointments and setbacks that science always
has in preparation for each one of us. It was
ominously Edgar Allan Poe who wrote the
following (abbreviated): “From childhood
hours I could not bring my passion from a
common spring”. My own outlandish passion
and curiosity for science has brought both
pros and cons to me. During my career I have
thus worked in many different disciplines,
including pathology, genetics, developmental biology, cell biology, evolution and so on.
Although each research interest usually had
to be built up from scratch, which slowed
down short-term progress, my search for
creative space on the interface between different research areas benefits from insights into
several different disciplines.
My first wobbling steps in the academic
world started with a BSc in Zoology and
Chemistry at Lund University. After my PhD
in Biochemistry in the stone ages, 1976, in
Lund, I went for postdoctoral research at the
National Institute of Medical Research in
Mill Hill, London. I returned to Sweden as
a postdoc at Uppsala University to work on
adenovirus mRNA processing. I took up my
first position as an independent scientist as a
guest professor at Umeå university. Against all
the Myc oncogene and cell proliferation in
odds, one of the most successful periods in my
normal cells1 and the first identification of a
scientific life was completed in a 4 m2 lab, sharnormal cell with autocrine growth control2.
ing desk with another scientist. This progress
From the discovery of genomic imprinting in
opened up an opportunity to take up a posihumans in 19933 to the discovery of systemic
tion at Karolinska Instiloss of imprinting in
tutet, first as a senior sciencolon cancer patients
“Of course we hope to
tist funded by the Swedish
in 19984, I have more
one day be able to use
Cancer Society and later as
recently established the
acquired information to
a professor funded by the
link between chromatin
not only diagnose early
Swedish Research Couninsulators and genomic
stages of cancer, but
cil for Natural Sciences.
imprinting5. Thus, the
also to design cancer
During this time, my intertargeted destruction of a
treatments.”
est in genomic imprinting
chromatin insulator at an
was initiated. After 16 years
imprinted locus perturbs
as a professor in Developmental biology at
stem cell populations in colon crypts to
Uppsala University and one wonderful year at
predispose to cancer6,7. This feature involves
MTC I am still on this track.
a novel epigenetic mark, poly(ADP-ribosyl)
Among my best results I would rate as
ation8, which has most recently been incorpobeing one of the first to make the link between
rated into the context of higher order chro-
If you want to make it
8
Department of Microbiology, Tumor and Cell Biology – MTC
matin conformation analyses 9,10,11. The most
recent work, since we came to MTC, places
such chromosomal networks in the context
of nuclear architecture and describes the
transfer of epigenetic states between chromosomes12,13,14.
Strong scientific competence at MTC
There are many reasons why I chose MTC
as my new address. Apart from wanting to
be part of the same institution that hosts a
legend, professor Georg Klein, I was attracted
by the strong scientific competence in cell
and molecular biology, particularly in cancer
pathways. The reason is that our work on
chromosomal networks point in these directions. I very much look forward to several
joint interactions with MTC scientists and
hope that MTC scientists will share my feeling for many interfaces of common interests.
Newcomers
We do have some know-how on chromatin,
epigenetics and nuclear architecture that
MTC scientists might be interested in.
Since my wonderful group moved to
MTC, we have further penetrated the network
of physically interacting chromosomes in
human stem cells as well as cancer cells. We
have uncovered links to cell signaling pathways and to apoptosis within these networks
and we are starting to think of our projects in
term of systems biology and nuclear architecture, which are two features not commonly
associated with each other. Our vision is not
only to understand epigenetics, genomics,
and nuclear architecture, I call this interface
“chromosomics”, but to take this interface
to stem/progenitor cells in particular and
explore the early stages of what can go wrong
in cancer. To this end, we have established a
large national and international network
of friends and colleagues who have complementing objectives. Of course we hope one
day to be able to use acquired information
not only to diagnose early stages of cancer, but
also to design cancer treatments.
Key publications
1. Pfeifer S, Goustin AS, Rydnert J, Wahlström T, Bjersing L and Ohlsson R.
Spatial and temporal pattern of cellular
myc oncogene expression in developing human placenta: implications for
embryonic cell proliferation. Cell 38,
585, 1984.
2. Goustin AS, Betsholtz C, Pfeifer S,
Persson H, Rydnert J, Bywater M,
Heldin C-H, Westermark B and Ohlsson R. Co-expression of the sis and
myc protooncogenes in early human
placenta suggests autocrine regulation
of trophoblast growth. Cell 41, 301,
1985.
3. Ohlsson R, Nyström A, Pfeifer-Ohlsson S, Hedborg F, Schofield P, Flam
F and Ekström T. IGF2 is parentally
imprinted during human embryogenesis and in the Beckwith-Wiedemann
syndrome. Nat Genet 4, 94, 1993.
4. Cui H, Horon IL, Ohlsson R, Hamilton
SR and Feinberg AP. Loss of imprinting in normal tissue of colorectal
cancer patients with microsatellite
instability. Nat Med 4, 1276, 1998.
The image shows a physical network of interacting chromatin regions from different chromosomes. The
data were generated from a library of interacting sequences in human embryoid bodies obtained by the
circular chromosome conformation capture method that the Ohlsson laboratory has invented. There is
a central node, representing a particular region on chromosome 2, which is connected to other nodes
and so-called outliers. We hypothesize that this network represents an overall regulatory feature of gene
expression that connects different modules of biological processes, such as different cell signaling pathways, to coordinate and diversify the expressivity of the genome.
5. Pant V, Mariano P, Kanduri C, Mattsson A, Lobanenkov V and Ohlsson R.
The nucleotides responsible for the
direct physical contact between the
chromatin insulator protein CTCF
and the H19 imprinting control region
manifest parent of origin-specific
long-distance insulation and methylation-free domains. Genes Dev 17, 586,
2003.
6. Sakatani T, Kaneda A, LacobuzioDonahue C, Carter M, Okano H, Ko
MS, Ohlsson R, Longo DL and Feinberg AP. LOI of Igf2 alters maturation
of intestinal epithelium and increases
adenoma formation in Min mice.
Science 307, 1976, 2005.
7. Feinberg A, Ohlsson R and Henikoff
S. The epigenetic progenitor origin of
human cancer. Nat Rev Genet 7, 21,
2006.
8. Yu WQ, Ginjala V, Pant V, Chernukhin
I, Whitehead J, Mukhopadhyay R,
Kanduri C, Oshimura M, Feinberg AP,
Lobanenkov V, Klenova E and Ohlsson
R. Poly(ADP-ribosyl)ation regulates
CTCF-dependent chromatin insulation. Nat Genet 36, 1105, 2004.
9. Göndör A and Ohlsson R. Transcription in the loop. Nat Genet 38, 1229,
2006.
10.Zhao Z, Tavoosidana G, Sjölinder M,
Göndör A, Mariano P. Wang S, Kanduri
C, Lezcano M, Singh Sandhu K, Singh
U, Pant V, Tiwari V, Kurukuti S and
Ohlsson R. Circular chromosomal
conformation capture (4C) uncovers epigenetically regulated networks
of intra- and interchromosomal
complexes. Nat Genet 38, 1341, 2006.
11.Ohlsson R. Widespread monoallelic
expression. Science 318, 1077, 2007.
12. Göndör A and Ohlsson R. Replication
timing and epigenetic reprogramming
of gene expression: a two-way relationship? Nat Rev Genet 10, 269, 2009.
13. Göndör A and Ohlsson R. Chromatin
crosstalk in three dimensions. Nature
461, 212, 2009.
14.Sandhu KS, Shi C, Sjölinder M, Zhao
Z, Göndör A, Liu L, Tiwari VK, Guibert
S, Emilsson L, Imreh MP and Ohlsson
R. Nonallelic transvection of multiple imprinted loci is organized by the
H19 imprinting control region during
germline development. Genes Dev 23,
2598, 2009.
Department of Microbiology, Tumor and Cell Biology – MTC
9
Newcomers
Sonia Laín
Bringing a Touch of
Chemistry to MTC
My research career started in 1988 in
Madrid where, after obtaining a University degree in biology, I carried out my PhD
thesis work on molecular biology aspects of
a poliovirus-like RNA virus infecting plants.
In 1992 I was awarded an EMBO fellowship
to work at the University of Gothenburg in
the Epstein-Barr Virus (EBV) field, a main
topic of research at the former Tumor Biology
department at the Karolinska Institutet (KI)
which is now part of MTC. Therefore, very
early on I became familiar with the exciting
EBV research by Georg and Eva Klein as well
as by other Tumor Biology members, many of
whom are currently at MTC.
After three years in Sweden, I moved to
Dundee in Scotland, where I began studies
on central tumor suppressor proteins such as
p53. Although I maintain a strong interest in
basic research, I quickly realized the importance of using the information and reagents
we were gathering for the discovery of therapeutics with antitumor activity. My interest
in using p53 to discover small molecules with
therapeutic potential began when I found the
Streptomyces metabolite leptomycin B to be
one of the most potent activators of p531 and
established the basis for a Phase I/II trial with
this compound through our interaction with
clinicians.
Fruitful and ongoing collaboration
My research on drug discovery was broadened when, together with professor David
Lane, one of the discoverers of p53 (and
adjunct Professor at MTC-KI) we obtained
substantial funding to perform compound
screens using a robust p53 activation cellbased assay2. More importantly, this work led
to a fruitful and ongoing collaboration with
the chemistry lab of Dr Nick Westwood in St
Andrews (Scotland).
By mid-2004 we had screened 34,000
compounds and identified a large number
of hits that we classified according to carefully chosen criteria. It was exciting to see
that two of these small molecules showed the
capacity to reduce tumor growth when tested
in animals3,4 as this validated our selection
procedures. New screens based on our protocols are currently underway. Another important aspect of this work was that we were
able to apply ground-breaking technologies
to solve the so-called “target identification
problem” of elucidating the mechanism of
action of small molecules in cells4. For example, we showed that one of our families of hit
molecules includes inhibitors of the protein
deacetylase activity of sirtuins, an extremely
interesting group of enzymes linking metabolism with gene expression. These enzymes
have been implicated in the ageing process
as well as in a variety of illnesses including
cancer and infectious diseases.
In the fall of 2008 I started a research lab
at MTC with five members so far constituting expertise in cell biology and chemistry.
Our plan is to continue with our approaches
for drug discovery focusing primarily on our
previously identified sirtuin inhibitors4, 5 as
well as on hit compounds from new screens.
Chemical biology is rapidly becoming popular in academia. Here, MTC can be
regarded as pioneering and I see, that there
are many opportunities for cross-fertilization
between research groups. We are keen to share
our best-characterised molecules, compound
collections and experience in chemical biology as well as to actively participate in training
in this highly exciting novel research area.
Our general goal is to develop well-characterised compounds that constitute high quality tools for research and/or potential therapeutics. This in-depth knowledge will help to
In the primary assay to screen for
compounds affecting p53 activity, a
p53 reporter mammalian cell line is
seeded into wells of 96 well plates
and each well is treated with a compound. Activation of p53’s transcriptional function is measured by colourimetric analysis of β-galactosidase
reporter activity expressed from the
p53-dependant lacZ gene.
10
Department of Microbiology, Tumor and Cell Biology – MTC
recognise clinical situations where they are
beneficial as well as to assess potential risks in
future clinical trials. Our hit compounds are
of interest for the treatment and understanding of diseases other than cancer and in this
regard, working at MTC opens up the possibility to test our hit compounds in collaboration with experts in a range of disease models.
Key publications
1. Laín S and Lane D. Improving cancer therapy by non-genotoxic activation of p53. Eur J Cancer 39, 1053,
2003.
2. Berkson R, Hollick J, Westwood N,
Woods J, Lane D and Laín S. A pilot
screening programme for small
molecule activators of p53. Int J
Cancer 115, 701, 2005.
3. Laín S, Hollick JJ, Campbell J, Staples
O, Higgins M, Aoubala M, McCarthy
A, Appleyard V, Murray KE, Thompson A, Mathers J, Holland SJ, Stark
MJR, Pass G, Woods J, Lane DP and
Westwood NJ. Discovery, in vivo
activity and mechanism of action of
a small-molecule p53 activator. Cancer Cell 13, 454, 2008.
4. Staples OD, Hollick JJ, Campbell J,
Higgins M, McCarthy A, Appleyard
V, Murray KE, Thompson A, Ronseaux S, Lane DP, Westwood NJ and
Laín S. Characterisation, chemical
optimisation and anti-tumor activity of tubulin poisons identified by
a p53-based phenotypic screen. Cell
Cycle 7, 3417, 2008.
5. Medda R, Russell RJM, Higgins M,
McCarthy AR, Campbell J, Slawin
AMZ, Lane DP, Laín S and Westwood NJ. Novel cambinol analogs
as sirtuin inhibitors: Synthesis,
biological evaluation and rationalization of activity. J Med Chem 52,
2673, 2009.
Newcomers
Lars-Gunnar Larsson
Tracking Down Fresh Footprints
of the Myc Tumor Protein
ers. Our expectations have been entirely
I graduated as a B. Sc. in Microbiology at
fulfilled since we moved to MTC in 2007.
Uppsala University 1982. However, since
From our side, we have contributed to further
then, I dedicated myself to Tumor Biology –
strengthen Tumor Biology research at MTC,
in my view a most fascinating field of research
in particular we concentrated on the analyembracing the most fundamental questions of
sis of different aspects of oncogene/tumor
how cell growth and development is regulated.
suppressor gene function and regulation. I
I got my PhD in Uppsala 1989 studying
presume my long experience in teaching is
oncogenes and their role in differentiation.
also an asset to the department.
During this period it became increasingly
clear that Myc, one the oncogenes I studied,
was a key player in tumor development. I
Visualize protein interactions
therefore decided to specialize in Myc and
In my career as an independent researcher I
made a postdoc 1991–94 in the laboratory of
have focused mainly on post-translational
Bernhard Lüscher in Hannover, Germany.
regulation of the oncoprotein/transcription
During this period I published one of the
factor Myc in response to signaling and tried
first papers on the Mad family of Myc antagoto elucidate the most important functions
nists1. By this time I had
of Myc. One of our major
achievements was the
become truly “addicted”
demonstration that phosto Myc, and returnphorylation of Myc reguing to Sweden in 1994 I
lates turnover by the ubiqstarted my own research
uitin/proteasome pathway
group at Rudbeck lab
and we subsequently idenin Uppsala continuing
tified the first E3 ubiquitin
Myc research. 1998 I got
Myc/Ras oncogene cooperativity in
ligase, Skp2, targeting Myc
a senior lecturer position
transformation. Apoptosis and cellular
for degradation2. Surprisat the Swedish University
senescence are two main barriers of
oncogenic transformation of cells. Lars- ingly, we found that Skp2
of Agricultural Sciences
son’s recent research suggests that
(SLU) in Uppsala and
simultaneously promotes
Myc and Ras complement each other
in 2001 I was appointed
Myc-driven
transcripby opposing each other’s anti-tumorigenic activities by repressing senesProfessor of Molecular
tion, thereby supporting
cence and apoptosis, respectively,
Genetics. I eventually
the so-called “Kamikazeresulting in tumor development.
decided to move on and
model” of transcription.
got recruited to MTC in 2007.
Another important finding was that Myc
The reason I chose MTC was its strong
directs transcription from all three RNA
position in Tumor Biology, and in particular
polymerases3. We also contributed to the
its long-standing and continuous interest in
developments of new methods to visualize
Myc and other transcription factors of releprotein interactions in living or fixed cells.
vance for Tumor Biology such as p53. Other
After moving to MTC we partially changed
reasons were my already ongoing collaboradirection and discovered that an important
tions with groups at MTC and other departfunction of Myc is the regulation of cellular
ments at the Karolinska Institutet (KI) and my
senescence4,5. Inspired by many other groups
increasing interest in translational research,
at MTC we have also put more emphasis
where MTC is strong. My expectations were
on translational research and are actively
to gain increased interactions, discussions
developing small molecules targeting Myc.
and collaborations with researchers sharFurther, we started research on Myc-driven
ing my interest in Tumor Biology. Further, I
tumor development in animal models. We
was looking forward that my research group
will develop these new aspects in the future
would become part of an exciting and interacin collaboration with other groups at MTC,
tive scientific environment, exchanging ideas
KI and in the world. I have also started to be
as well as technical know-how through semiengaged more actively in teaching at basic and
nars and daily contacts with other researchadvanced courses at MTC and KI and hope to
be able to contribute more to teaching activities in the future.
Key publications
1. Larsson L-G, Pettersson M, Öberg F,
Nilsson K and Lüscher B. Expression
of mad, mxi1, max and c-myc during
induced differentiation of hematopoietic cells: opposite regulation of
mad and c-myc. Oncogene 9, 1247,
1994.
2. von der Lehr N, Johansson S, Wu S,
Bahram F, Castell A, Cetinkaya C,
Hydbring P, Weidung I, Nakayama
K, Nakayama KI, Söderberg O, Kerppola K and Larsson L-G. The F-box
protein Skp2 participates in c-Myc
proteosomal degradation and acts as
a cofactor for c-myc-regulated transcription. Mol Cell 11, 1189, 2003.
3. Arabi A, Wu S, Ridderstråle K,
Bierhoff H, Shiue C, Fatyol K,
Fahlén S, Hydbring P, Söderberg
O, Grummt I, Larsson L-G and
Wright APH. c-Myc associates with
ribosomal DNA and activates RNA
polymerase I transcription. Nat Cell
Biol 7, 303, 2005.
4. Hydbring P, Bahram F, Su Y, Tronnersjö S, Högstrand K, von der Lehr
N, Lilischkis R, Hein, Wu S, Vervoorts
J, Henriksson M, Grandien A, Lüscher
B and Larsson L-G. Phosphorylation
by Cdk2 is required for Myc to repress
Ras-induced senescence in cotransformation. Proc Natl Acad Sci USA
107, 58, 2010.
5. Campaner S, Doni M, Hydbring P,
Verrecchia A, Bianchi L, Sardella D,
Schleker T, Perna D, Tronnersjö S,
Murga M, Fernandez-Capetillo O,
Barbacid M, Larsson L-G and Amati
B. Cdk2 suppresses cellular senescence induced by the myc oncogene.
Nat Cell Biol 12, 54, 2010.
Department of Microbiology, Tumor and Cell Biology – MTC
11
Newcomers
Pontus Aspenström
The Secret Garden
of Rho GTPases
with the
clear objective to become a botanist. I wanted
to spend my life knowing all about flowers; it
was not on the agenda to lock myself up within
the confinement of a laboratory. Strangely
enough, this was exactly what I did as a PhD
student when I found myself studying microfilament proteins, such as actin. The concept
that microfilaments acted as the primum
movens for cell migration was gaining general
acknowledgement during the late eighties.
However, the signaling pathways that regulate
the dynamic reorganization of the actin filament system were essentially unknown.
I started my academic studies
Wiskott-Aldrich syndrome protein
The identification of the Ras-like small
GTPases Rac and Rho as the key elements in
the signaling pathways that link transmembrane receptors and the actin filament system
resulted in a true shift of paradigm. This seminal discovery by Alan Hall and co-workers
triggered my interest in Rho GTPases and I
decided for a post-doctoral visit to his laboratory at University College London. There,
I became engaged in the hunt for binding
partners for the Rho member Cdc42. During
this period, we identified the Wiskott-Aldrich
syndrome protein (WASP)1, a key component
in the regulation of actin polymerization and
Rho GTPases control the integrity and reorganization of the actin filament system. Expression of
the Rho member RhoD (green) in a porcine aortic
endothelial cell triggers the formation of actin bundles (red), as well as, finger-like protrusions, so
called filopodia, at the cell periphery.
12
Department of Microbiology, Tumor and Cell Biology – MTC
PAR-6, a component of the polarity complex
involved in the regulation of directed cell
migration as well as for the establishment of
epithelial cell polarity2.
I was also involved in the identification
of F-BAR domain-containing proteins3. The
function of the F-BAR proteins is to coordinate cytoskeletal organization and membrane
dynamics as they can bind and induce curvature to lipid bilayers.
A new family of GTPases
After returning to Sweden, I became a group
leader at the Ludwig Institute for Cancer
Research in Uppsala and my group continued
to study the link between Rho GTPases and
cytoskeletal organization4. In the search for
novel Rho GTPases, a new family of GTPases
was identified, which we named Miro (mitochondrial Rho)5. The Miro GTPases have
emerged as key components in the regulation of mitochondrial movement along
microtubules. Deregulated mitochondrial
transport and morphology is linked to severe
neuropathogical diseases, such as type 2A
Charcot-Marie-Tooth disease and dominant
optic atrophy.
After twelve years in Uppsala, the time was
ripe to move on and I settled with my group
at MTC during 2008. My main reason for this
decision was the feeling that this department
could provide a dynamic and attractive work
environment. Our work is to a large extent
based on microscopy and the presence of the
KI visualization facility (KIVIF) at MTC will
be an asset. We intend to continue to study
Miro, as well as the less known members of
the Rho GTPases, to identify novel binding
partners and to study how these networks of
interactions can orchestrate cell functions.
The microenvironment in tumors differs
substantially from the surrounding tissue
with regard to cell stiffness. Therefore, we
want to study how extracellular force and
rigidity affect vital processes such as cell
contraction, migration and cell cycle progression. The diverse environments of the Karolinska Institute and the proximity to the Karolinska University hospital will be a benefit
since the emerging view strongly indicates
that Miro and Rho GTPases, as well as several
of their binding partners, are dysfunctional or
deregulated in cancer and in neuronal pathologies. It is my hope to increase the connections to clinical expertise in order to define to
what extent the main targets for our research
interests could also serve as targets for diagnostics and medication in human disease.
Key publications
1. Aspenström P, Lindberg U and
Hall A. Two GTPases, Cdc42 and
Rac, bind directly to a protein
implicated in the immunodeficiency disorder Wiskott-Aldrich
Syndrome. Curr Biol 6, 70, 1996.
2. Johansson A-S, Driessens M and
Aspenström P. The mammalian
homologue for the Caenorhabditis elegans polarity protein PAR-6
is a binding partner for Cdc42. J
Cell Sci 113, 3267, 2000.
3. Aspenström P. A Cdc42 target
protein with homology to the
non-kinase domain of FER has
a potential role in regulating the
actin cytoskeleton. Curr Biol 7,
479, 1997.
4. Aspenström P, Fransson Å and
Saras J. The Rho GTPases have
diverse effects on the organization of the actin filament system.
Biochem J 377, 327, 2004.
5. Fransson Å, Ruusala A and
Aspen­
ström P. Atypical Rho
GTPases have roles in mitochondrial homeostasis and apoptosis.
J Biol Chem 278, 6495, 2003.
Department of Microbiology, Tumor and Cell Biology – MTC
13
Awards, Major Grants and Appointments
2007
n Foreign Adjunct Professors
Alexander von Gabain, Intercell AG, Vienna
Biocenter, Austria; Margaret Liu, Carolina Vaccine Institute, University of North Carolina,
USA; Birgit Lane, University of Dundee, Scotland UK and Centre for Molecular Medicine,
Singapore, and Sir David Lane, University of
Dundee, Scotland UK and Institute of Molecular and Cell Biology, Singapore, were appointed
Foreign Adjunct Professors at MTC.
n Adjunct Professors
Annelie Brauner, Anneka Ehrnst, Gunilla
Källenius and Åke Lundkvist were appointed
Adjunct Professors at MTC.
n New Professor
Agneta Richter-Dahlfors was appointed
Professor in Cellular Microbiology.
n From MTC to the Government
Former MTC group leader and Associate
Professor Ewa Björling was appointed Minister of Foreign Trade in the Swedish Government by Prime Minister Fredrik Reinfeldt.
n Peoples Republic of China National
Friendship Award
Ingemar Ernberg received the People’s Republic of China National Friendship Award
from the president of the People’s Republic
of China, Hu Jintao, and the premier of the
People’s Republic of China, Wenjao Bao, in
Beijing, China. The Award is the most important Chinese prize to foreign experts.
n Peoples Republic of China Award for
International Cooperation in Science and
Technology
Ingemar Ernberg received the People’s Republic of China Award for International Cooperation in Science and Technology by the
Ministry of Science and Technology (MOST)
in Beijing.
n Best Scientific Achievement of the year
Annelie Brauner was awarded Best Scientific
Achievement of the year at the Karolinska
University Hospital, Solna.
14
Department of Microbiology, Tumor and Cell Biology – MTC
n Sven Gard Scholarship
Hanna Sjölin received the Sven Gard Scholar­
ship for the best Virology Thesis in 2006.
n New Associate Professors (Docents)
Maria Isaguliants and Tobias Allander were
appointed associate professor at KI.
n Honorary Member of the Slovak Pediatric
Society
Annelie Brauner was elected Honorary
Member of the Slovak Pediatric Society, a lifelong membership.
n Knut and Alice Wallenberg Foundation
Klas Kärre and co-applicants received a grant
for the purchase of state of the art FACS
equipment.
n Ministry of Education of China
n MTCers of 2007
The organizers of the MTC pub: “The Muppet
Crew”, Danika Shepsis, Emma Hutric and
Alberto Cagigi.
n MTC Scientist of 2007
Mats Wahlgren.
n MTC Paper of 2007
From the Yihai Cao group: “Angiogenic
factors FGF2 and PDGF-BB synergistically
promote murine tumor neovascularization
and metastasis” Lars Johan Nissen, Renhai
Cao, Eva-Maria Hedlund, Zongwei Wang,
Xing Zhao, Daniel Wetterskog, Keiko Funa,
Ebba Bråkenhielm and Yihai Cao.
J Clin Invest 117, 2766, 2007.
n MTC Pedagogical Prize of 2007
Roland Möllby.
Hao Mo received the Prize for Outstanding
Chinese PhD Students, which comes with an
award from the Chinese government.
n Positions from the Swedish Research
Council
Pontus Aspenström received a 6-year Senior
Researcher position for molecular studies of
cell motility.
Laura Plant and Markus Sköld received
4-year Research Assistant positions.
n Position from the Swedish
Childhood Cancer F
­ oundation
Marina Vita received a
2-year postdoc position.
n Position from the
Swedish Cancer S­ ociety
Marie Arsenian Henriksson received the Senior Investigator
Award from the Swedish Cancer
Society.
n MTC Annual Talks Poster prizes
2007
Keira Melican, Emilie Flaberg and
Jakob Lovén.
Ingemar Ernberg,
recipient of the ­Peoples
Republic of China
National Friendship
Award.
2008
n New Professors
Rolf Ohlsson was appointed a strategic professorship in Genomic Integrity.
Birgitta H
­ enriques Normark was appointed
Professor in Medical Microbial Pathogenesis.
n KI University Board
Klas Kärre was elected member of the University Board of the Karolinska Institutet for the
coming three years.
n KI Pedagogical Prize
Annelie Brauner and Tomas Cronholm (MBB)
received the KI Pedagogical Prize. Annelie
Brauner received the prize for her outstanding
achievements in education.
n Science Business Innovation Board
Hans Wigzell received The Bridge Award
“for an individual who has done the most
to promote policies for entrepreneurship in
university or public research institutions”.
n Chairman of the KI Cultural Board
Ingemar Enberg was elected chairman of the
KI Cultural Board, replacing professor Jan
Lindsten.
n Herbert J. Block Memorial Lectureship
Award
Georg Klein received the Herbert J. Block
Memorial Lectureship Award, Ohio State
University, USA.
n Bill and Melinda Gates Foundation
Gunilla Karlsson Hedestam and Bruce Beutler
at the Scripps Research Institute received
a 3-year grant to study “Immunoadjuvant
effects influencing induction of anti-viral B
cell responses”.
n Torsten och Ragnar Söderberg Foundation
Sven Pettersson and former MTC PhD student
Fredrik Bäckhed received a grant for the
project “Gut flora, peroxisome proliferating activating receptor (PPARg), adipocytes,
macrophages and mechanisms underlying
obesity”.
Yihai Cao received a grant for the project
“Angiogenesis and lymphangiogenesis in
cancer and metastasis”.
Galina Selivanova recieved a grant for the
project “Targeting p53 to combat cancer:
From laboratory bench to patient”.
Poster competition winners at the MTC Annual Talks 2007 from left to right, PhD students Keira Melican,
Emilie Flaberg and Jakob Lovén.
n Positions from the Swedish Research
­Council
Martin Rottenberg received a 6-year Senior
Researcher position for studies of congenital
immunity.
Birgitta Henriques-Normark received a 50
percent 6-year Senior Researcher position in
clinical bacteriology.
Bence Rethi and Björn Önfelt received 4-year
Research Assistant positions.
Hanna Sjölin received a 3-year postdoc
fellowship.
Katarina Reis received a 2-year postdoc position.
n KI Senior Researcher
The Board of Research at Karolinska Institutet
granted Katrin Pütsep funds for employment
as a Senior Researcher for two years.
n KI Ethics Council
Marie Arsenian Henriksson was appointed as
member of the KI Ethics Council.
n Jonas Söderqvists Scholarship
Åsa Hidmark received the 2008 year scholar­
ship from Jonas Söderqvists Scholarship
Foundation.
n The Swedish Foundation for Strategic
Research
Björn Önfelt was awarded a grant within the
program “Future Research Leaders” to work at
MTC and the Department of Applied Physics,
The Royal Institute of Technology (KTH).
n Wenner-Gren Foundations fellowship
Hanna Sjölin has been awarded a WennerGren Fellowship for 3 years to study in the US
with the possibility for an additional two year
Research Assistant position on returning to
Sweden.
n MTC “15 year Celebration” Poster prizes
Yuan Xue, Sandra Nilsson and Petra Luthje.
n MTCers of 2008
The Economy Unit: Lada Larsson, Carina
Lingonbacke and Eva Wichert.
n MTC Junior Scientist of 2008
Anna Nilsson.
n MTC Paper of 2008
From the Petter Höglund group: “The strength
of inhibitory input during education quantitatively tunes the functional responsiveness of
individual natural killer cells.” Petter Brodin,
Tadepally Lakshimikanth, Susanne Johansson,
Klas Kärre and Petter Höglund. Blood 113,
2434, 2009.
n MTC Photographer of 2008
Lennart Nilsson.
n MTC Artist of 2008
Ewert Linder.
n MTC Life Science Achievement Award
Hans Wigzell.
n The Swedish Childhood Cancer
Foundation
Helén Nilsson received a 2-year postdoc position.
Department of Microbiology, Tumor and Cell Biology – MTC
15
2009
n KI Distinguished Professor Awards
Yihai Cao, Klas Kärre, Staffan Normark and
Mats Wahlgren received this special grants
for KI professors who carry out research of
the highest quality at the international level.
n Torsten och Ragnar Söderberg Foundation
Mats Wahlgren received the 5-year Söderberg
Research Professorship in Medicine awarded
by The Royal Swedish Academy for Sciences.
n The Royal Academy for Engineering
Science (IVA)
Hans Wigzell was awarded the Gold Medal for
his entrepreneurial work within the field of
Biotechnology.
n EU grant
Martin Rottenberg received a 3-year grant
to coordinate an EU consortium, including
nine countries, to study “Mycobacterium-host
interactions”.
n Position from Marianne och Marcus
Wallenbergs Foundation
Petter Höglund received a 3-year position.
n International AIDS Vaccine Initiative
Gunilla Karlsson Hedestam received a 5-year
grant to investigate B cell responses to the
HIV-1 envelope glycoproteins with a focus on
induction of broadly neutralizing antibodies.
n The Heart and Lung Foundation and King
n The Royal Swedish Academy of Sciences
Klas Kärre was elected member of the Royal
Swedish Academy of Sciences.
Oscar II’s Jubilee Foundation
Markus Maeurer, Martin Rottenberg and
Markus Sköld were awarded a grant for
research on a recombinant BCG vaccine
expressing novel antigens.
n Academic Medal
n Position from the Swedish Research
Georg Klein was awarded the medal of
Biomedicum from Helsinki, Finland.
Council
Gunilla Karlsson Hedestam was awarded
a 6-year Senior Research position for
development of novel innovative vaccines.
n Research Strategy Committee (FSK)
Galina Selivanova and Sten Nilsson were
appoined coordinators for a new translational research center entitled: “Preclinical
and clinical development of novel targeted
anti-cancer therapies (ACT!).” They received
the grant for the initial period 2010-2012 by
the Research Strategy Committee, which is
a collaborative committee between KI and
Stockholm County Council (SLL).
n Position from the Swedish Cancer Society
Petter Höglund received a 50 percent 6-year
research position from the Swedish Cancer
Society.
n Bill and Melinda Gates Foundation
Markus Maeurer received a 2-year grant
to study “Pattern recognition in immune
responses directed against Mycobacterium
tuberculosis (Mtb) using high content peptide
microarray chips”.
n New Associate Professors (Docents)
Annika Karlsson and Elena Kasuba were
appointed associate professor at KI.
n The Swedish Society for Medical Research
(SSMF)
Mattias Forsell received a postdoctoral scholarship.
n MTCer of of 2009
John Sennett.
n MTC Scientists of of 2009
Galina Selivanova and Yihao Cao.
n MTC Paper of of 2009
From the Rolf Ohlsson group: ”Nonallelic
transvection of multiple imprinted loci is
organized by the H19 imprinting control
region during germline development”. Kuljeet
Singh Sandhu, Chengxi Shi, Mikael Sjölinder,
Zhihu Zhao, Anita Göndör, Liang Liu, Vijay K
Tiwari, Sylvain Guibert, Lina Emilsson, Marta
P Imreh and Rolf Ohlsson. Gen Dev, 23, 2598,
2009.
n MTC Pedagogical Prize of 2009
Anneka Ehrnst.
Markus Maeurer
briefing Bill Gates on
drug resistance in
tuberculosis.
Photo: Ewert Linder
16
Department of Microbiology, Tumor and Cell Biology – MTC
Impressions from the MTC 15 years anniversary
Speakers at the MTC Jubilee Symposium at Nobel forum June 4-5, 2008.
Georg Klein explaining the main principles of tumor biology to an interested audience of technical and administrative staff in June 2008.
KI President Harriet Wallberg-Henriksson opens the Jubilee Symposium on
June 4, 2008.
Hannah Akuffo, Francesca Chiodi and Peter Krammer mingling during a coffee
break between the lectures on the Jubilee Symposium focusing on TB and HIV
in November 2008.
Department of Microbiology, Tumor and Cell Biology – MTC
17
Scientists in Focus
Birgitta Henriques Normark
From the Clinics
to Basic Science
I received my medical degree from the
Karolinska Institutet (KI) in 1983 and then
worked as a clinician for several years at
different hospitals in the Stockholm area. I
developed my interest in respiratory tract
infections when I worked as an ear-noseand throat doctor/surgeon for a long time at
Huddinge hospital. I first thought I would
become a clinical fellow but when, before I did
my internship, I was involved in research on
the bacterium Clostridium difficile together
with professor Monica Thelestam at the
Department of Bacteriology. I enjoyed it
so much that I decided that I would like to
continue with research on microbes.
After internship at Huddinge and
Södertälje hospitals I started as a physician
at The National Bacteriological Laboratory
(SBL) that later became The Swedish Institute for Infectious Disease Control (SMI). I
defended my thesis in the year 2000, became
docent in 2004 and professor at KI in 2008.
Translational approach
My research focuses on the major pathogen
Streptococcus pneumoniae (or pneumococci)
and has a translational approach going from
the patient to the small molecule. Pneumococci are a common cause of morbidity and
mortality world-wide, primarily among
children and the elderly, but also a common
colonizer of the nasopharynx of pre-school
children. To understand why pneumococcal
Electron micrograph of pneumococcal pili.
18
Department of Microbiology, Tumor and Cell Biology – MTC
infection can result either in healthy carriage,
local or invasive disease we first built up a
molecular epidemiological platform allowing us not only to serotype but also to group
pneumococcal isolates into genetically related
clonal types. We identified pneumococcal
clones with different likelihood of causing
carriage and invasive disease respectively.
We also began to understand the correlation between clonal types and the severity of disease and other clinical aspects
such as disease type, mortality and to what
extent underlying diseases in the patients
are required for certain clonal types to cause
disease. Isolates belonging to defined clonal
types were characterized in mice model
infection and a genome wide analysis was
performed by microarray analyses using a
whole genome array that we set up, as well
as whole genome sequencing. We identified
pathogenicity islets being of importance
for pneumococcal epidemicity as well as for
disease outcome. A major finding was that
pneumococci carry pilus like structures on
their surface.
a special emphasis on cell wall recognition by
NOD proteins and the potential immunomodulatory role played by peptidoglycan
receptors.
Our research program also embraces other
pathogens such as Group A streptococci,
enterococci and Chlamydia.
Promotes global spread
5 selected references
The biogenesis, regulation, epidemiology, and
structure as well as the role of the pilus and its
components in disease are now being actively
pursued. Our recent finding that piliation
promotes global spread of antibiotic resistant pneumocococcal clones is particularly
important and suggests that pilus
proteins may be used as protective
antigens in vaccines against the
spread of resistant pneumococci.
Host defense factors and innate
immune responses involved in
pneumococcal pathogenesis are
actively being pursued in vivo and
in vitro. Thus, we have defined the
role of Toll like receptor signaling
in host defense, characterized the
role of neutrophil extracellular traps
(NETs) and the interaction with
dendritic cells in pneumococcal
infection. The role of various host
components in clearing the organism at local sites, and in the blood
stream, is being pursued as well with
1. Beiter K et al., An endonuclease
allows Streptococcus pneumoniae
to escape Neutrophil Extracellular
Traps. Curr Biol 16, 1, 2006.
2. Barocchi M et al., A Pneumococcal
pilus influences virulence and host
inflammatory responses. Proc.
Natl Acad Sci USA 103, 2857, 2006.
3. Muschiol S et al., A small molecule inhibitor of type III secretion inhibits different stages of
the infectious cycle of Chlamydia
trachomatis. Proc Natl Acad Sci
USA 103, 14566, 2006.
4. Sjöström K et al., Clonal success of
piliated penicillin non-susceptible
pneumococci. Proc Natl Acad Sci
USA 104, 12907, 2007.
5. Henriques-Normark B et al., The
rise and fall of bacterial clones:
with a focus on Streptococcus
pneumoniae. Nat Microbiol Rev 6,
827, 2008.
Scientists in Focus
Hannah Akuffo
Towards an Adjunct
Professorship in Parasitology
I was educated in Ghana and England and
have been keen to explore different areas
within my life-long education. Bachelors’
degree in Biochemistry and Food Science in
Ghana made me curious about immunology.
Master of Science in Immunology in London
focusing on BCG infection in mice, created
an interest in a mouse model of leprosy. PhD
and postdoctoral studies in London on the
mechanisms of resistance to Mycobacterium
lepraemurium infection in mice as a model
for leprosy, led to a decision to study human
leprosy. A position as senior scientist in Ethiopia taught me a lot about this disease, but I
was also introduced to leishmaniasis, which
like leprosy presents a spectrum of clinical
manifestations.
Coming to Sweden (for love) in 1986 and
working at the Karolinska Institutet, but now
a dedicated leishmaniac, my work concentrated on finding out why Swedish healthy
individuals, unexposed to Leishmania, can
still have a response to Leishmania parasites.
The answer pointed to natural killer (NK)
cells, taking my research interest towards
understanding the interaction between NK
cells and Leishmania parasites and the role of
NK cells in the development and outcome of
Leishmania infection. Working with human
leishmaniasis has resulted in collaborations
in Ethiopia, Iran, Nicaragua, Somalia, Sudan,
USA and Uzbekistan with a coupled interest
Ulcer formation during Leishmania major induced
cutaneous leishmaniasis is associated with the
expression of TRAIL and Fas in the epidermis.
Photo by Liv Eidsmo.
in research training of students from many of
these countries.
The irony for me and my heritage is that
leishmaniasis is one of the few parasitic
diseases that was not found in Ghana in the
1990s, which drew me to explore another
disease which is manifested as a clinical spectrum, namely river blindness caused by the
worm Onchocerca volvulus. These cellular
immunology studies were performed in the
Volta region of Ghana, in the World Health
Organization (WHO) dedicated clinical trials
center in Hohoe. The possibility to engage
my students from Guatemala and Sweden in
these activities in Ghana was very enriching.
In 2003, an outbreak of skin ulcers in the Volta
region of Ghana was found to be caused by
leishmaniasis!
Most people would advice that to get on
in science one has to focus. I, however, believe
that the knowledge gained from immunological studies on Mycobacteria, Leishmania
and worms have culminated in our studies
on de-worming and improving vaccine efficacy. Linking some experience from antileishmaniasis vaccine research in dogs, we
set out to study the effect of de-worming on
the response to BCG, the anti-tuberculosis
vaccine which is highly effective in high
income countries, but less effective in low
income countries. Our studies in humans
showed that de-worming indeed does
improve BCG response and studies in mice
showed that worms certainly compromised
the efficacy of BCG vaccine against Mycobacterium tuberculosis infection.
I had the good fortune, at the end of
1998, to be employed by The Department
of Research Cooperation, SAREC, at the
Swedish International Development Agency
(Sida). This led me to obtain an increased
understanding of research capacity building.
In 2006 I formally became an Adjunct Professor in Parasitology.
I am in a truly privileged situation. I have
been able to remain in the academia doing
research on diseases of importance for low
income countries and at the same time I
am able to work with Sida to contribute to
enhancing the conditions for research and
research capacity in these countries. This
work also allows me to be involved in changes
at the institutional and national levels to
foster research efforts by individuals in low
income countries. The experience from my
work at MTC feeds directly into my work at
Sida and vice versa.
5 selected references
1. Eidsmo L et al., The contribution
of the Fas/FasL apoptotic pathway
in ulcer formation during Leishmania major-induced cutaneous
Leishmaniasis. Am J Pathol 166,
1099, 2005.
2. Nylén S et al., Surrogate markers
of immunity to Leishmania major
in leishmanin skin test negative
individuals from an endemic area
re-visited. Vaccine 24, 6944, 2006.
3. Lieke T et al., Leishmania surface protein gp63 binds directly
to human natural killer cells and
inhibits proliferation. Clin Exp
Immunol 153, 221, 2008.
4. Elias D et al., Poor immunogenicity of BCG in helminth infected
population is associated with
increased in vitro TGF-beta production. Vaccine. 26, 3897, 2008.
5. Nylén S and Akuffo H. Tracing
immunity to human leishmaniasis. Future Microbiol 4, 241, 2009.
Department of Microbiology, Tumor and Cell Biology – MTC
19
Hans G Boman in Memoriam
(1924–2008)
Professor Hans G Boman
was an exceptionally creative
scientist whose ground-breaking discoveries have changed
our understanding of the firstline of immune defence. He
initiated the flourishing field of
innate immunity and antimicrobial peptides. As professor
emeritus, he moved from Stockholm University to join MTC
and Karolinska Institutet (KI) in
1997. During his time at MTC,
Hans G Boman was awarded
some of the most important
Nordic prizes, the Nordic Fernström Prize (2000), the Söderberg Prize (2002) and the Olof
Rudbeck Prize (2005). He was
honorary Doctor in Medicine at
Umeå University and a member
of the Royal Academy of Sciencies in Sweden. Hans is deeply missed by his
colleagues and friends (Stromminger, 2009;
Pütsep and Faye, 2009).
peptide production is developmentally regulated rather than regulated by environmental
challenge (Pütsep et al., 2000).
One third of the world population is colonized
by the gastric bacterium Helicobacter
Hans G Boman’s research at MTC
pylori,
but most infections are asymptomatic.
At MTC Hans formed, together with his
Together with his colleagues at MTC, Hans
wife Anita, what became his last research
discovered that this commensal bacterium
group, in which he focused on antimicropossess antibacterial activity, but is selfbial peptide expression and its regulation in
resistant. Hans, and others, have suggested
animals and man. As animal model systems,
that H. pylori may have beneficial effects on
Hans used frogs and mice. Frogs produce
its host, as this antimicrobial peptide may
high amounts of antimicrobial peptides in
contribute to the defence against gastrointheir skin secretions. In a series of elegant
testinal pathogens (Pütsep et
experiments in collabo“Hans G Boman’s
al., 1999).
ration with colleagues in
intellectual insight
Working at a medical
Rome, Hans could demonreached far beyond
university, Hans wished to
strate that glucocorticoids,
the scientific world.”
identify diseases with abera class of steroid hormones,
rant expression of antimiinhibit antimicrobial peptide
crobial peptides. Indeed, in a Lancet publiexpression in frog skin resulting in higher
cation from 2002, Hans’ research group, in
bacterial load (Miele et al., 1998; Simmaco et
collaboration with haematologists at Karoal., 1998). Mice, in turn, protect the epithelinska University Hospital, demonstrated
lial integrity of the small intestine largely
deficiency in the production of the antimithrough high production of antimicrobial
crobial peptide hCAP18/LL-37 in neutrophils
peptides. Unexpectedly, germ-free animals
of subjects with severe congenital neutropeand conventional mice showed essentially
nia (Kostmann disease) (Pütsep et al., 2002).
very small differences in peptide production.
Kostmann subjects have an abnormally low
Thus mice are already at birth prepared for a
number of neutrophils, the most important
co-existence with microbes as antimicrobial
20
Department of Microbiology, Tumor and Cell Biology – MTC
type of white blood cell, in
their blood. The loss of LL-37
production in neutrophils is
most likely one reason for the
subjects still being infection
prone, in spite of treatment with
human granulocyte colonystimulating
factor, which
elevates neutrophil levels. Since
that publication a number of
other groups have demonstrated the importance of antimicrobial peptides in various
diseases, such as inflammatory
bowels diseases, urinary tract
infections and skin disorders.
Foundation of a novel
research field
Hans entered his scientific
career with the perspective
of a biochemist. He received
his PhD in Biochemistry in 1958 under the
guidance of Arne Tiselius at Uppsala University, followed by post-doctoral studies on
protein biosynthesis in bacteria in the excellent and dynamic research environment of
Fritz Lipmann, a former Nobel Prize laureate, at the Rockefeller Institute in New York.
He returned to Uppsala University in 1960,
where he established his research group at the
Department of Biochemistry with focus on
the biosynthesis of tRNAs.
In 1966, Hans was appointed the first
professorship in Microbiology ever in Sweden
at Umeå University. Initially, he focused on
mechanisms of bacterial resistance against
penicillins. However, he was soon fascinated
by the discrepancy between the mounting time for an efficient adaptive immune
response and the fast growth of bacteria.
There had to be other efficient mechanisms
for protection! Since insects lack the key
factors of adaptive immunity, B cells and T
cells, Hans turned to insects as model systems.
In 1972 Hans demonstrated, in collaboration with Bertil Rasmusson, the existence of
an inducible defence system in Drosophila
(Boman et al., 1976).
When Hans later moved to Stockholm
University he devoted his research to unrav-
elling the characteristics and mechanisms of
this inducible defence system. It took Hans
almost a decade and a change to the silk moth
Hyalophyra cecropia as a model system to
demonstrate that peptides with antimicrobial
activity mediated the previously described
inducible defence (Steiner et al., 1981).
Other research groups entered the field
and soon it became evident that immune
defence mediated by antimicrobial peptides
is an ancient and evolutionary conserved
principle throughout all kingdoms of life. In
collaboration with Viktor Mutt at KI, Hans
characterized the antimicrobial peptides
PR-39 and NK-lysin. The cloning of PR-39 by
Hans and colleagues lead to the discovery of
a major human antimicrobial peptide, LL-37
(Agerberth et al., 1995).
New scientific avenues
Throughout his scientific life, Hans strived to
find new scientific avenues by asking the most
simple, but unanswered, questions. Hans
communicated this philosophy and his strategy for performing good research in an essay –
How do you do Life Science? – which is available at the KI-website (Hans G Boman, KI
website). Hans was a modern and free-thinking supervisor who took great interest and
pleasure in discussions with students. He was
an appreciated lecturer and when he moved
to KI he engaged in graduate student courses,
teaching theoretical research methodology,
References
l
l
l
l
Boman H, Nilsson GI and Rasmusson B. Inducible antibacterial defence
system in Drosophila. Nature 237, 232,
1976.
Steiner H, Hultmark D, Engström A,
Bennich H and Boman HG. Sequence
and specificity of two antibacterial
proteins involved in insect immunity.
Nature 292, 246, 1981.
Agerberth B, Gunne H, Odeberg J,
Kogner P, Boman HG and Gudmundsson GH. FALL-39, a putative human
peptide antibiotic, is cysteine-free and
expressed in bone marrow and testis.
Proc Natl Acad Sci USA 92, 195, 1995.
Miele R, Ponti D, Boman HG, Barra D
and Simmaco M. Molecular cloning of
a bombinin gene from Bombina orientalis: detection of NF-κB and NF-IL6
binding sites in its promoter. FEBS Lett
431, 23, 1998.
The antimicrobial peptide Cecropin P1 was previously considered to be synthesized by the pig intestine.
Hans and his collegues could demonstrate that contaminating nematodes were the source of Cecropin
P1 (Andersson et al., 2003).
The large parasitic nematode worm of the genus Ascaris infects human (Ascaris suum) and pig
(Ascaris lunbricoides) intestines. Mass spectrometry analysis of A) HPLC fraction with antimicrobial activity from Ascaris lumbricoides B) synthetic antimicrobial peptide Cecropin P1.
Hans G Boman welcomed state-of-the-art techniques and with financial support from the Wallenberg
foundation, he brought a MALDI-TOF spectrometer to MTC, which is still in use.
The art of science as decision making. Being a
scientist driven by curiosity, he was a frequent
visitor of seminars on virtually any topic,
including the MTC student seminars.
In 2003, when Hans’ physical health
began to create hindrances, he declined the
position as a group leader. He was however,
still actively participating in the scientific
discussions and he concluded his view and
perspectives on peptide-mediated defense
in a review (Boman, 2003) and on science
and academia in an autobiograpy (Boman,
2002). Hans inspired many colleagues at MTC
and elsewhere at KI to search deeper into
l
l
l
l
Simmaco M, Mangoni ML, Boman A,
Barra D and Boman HG. Experimental infections of Rana esculenta with
Aeromonas hydrophila: a molecular mechanism for the control of the
normal flora. Scand J Immunol 48, 357,
1998.
Pütsep K, Brändén CI, Boman HG and
Normark S. Antibacterial peptide from
Helicobacter pylori. Nature 398, 671,
1999.
Pütsep K, Axelsson LG, Boman A,
Midtvedt T, Normark S, Boman HG
and Andersson M. Germ-free and colonized mice generate the same products
from enteric prodefensins. J Biol Chem
275, 40478, 2000.
Pütsep K, Carlsson G, Boman HG and
Andersson M. Deficiency of antibacterial peptides in patients with morbus
Kostmann: an observation study.
Lancet 360, 1144, 2002.
the field of innate immunity. In addition to
being an enthusiastic scientist, he surprised
his colleagues with an arts exhibition of his
collage paintings at KI in 2006.
Hans G Boman’s intellectual insight
reached far beyond the scientific world. It was
genuinely enjoyable to discuss with him any
subject, may it be movies, politics, literature,
arts or music. Every summer, Hans together
with his wife Anita went sailing for at least six
weeks. Here, Hans could charge his batteries
and find new perspectives on unsolved questions in biology.
Katrin Pütsep
l
l
l
l
l
Boman HG. En egensinnig biologs liv,
skildrat av honom själv. In free translation: ’The life of a headstrong biologist,
told by himself.’ Autobiography 2002.
Boman HG. Antibacterial peptides:
basic facts and emerging concepts. J
Intern Med 254, 197, 2003.
Strominger JL. Pillars of Immunology.
Animal antimicrobial peptides: ancient
players in innate immunity. J Immunol
182, 6633, 2009.
Pütsep K and Faye I. Hans G Boman
(1924-2008): pioneer in peptide-mediated innate immune defence. Scand J
Immunol 70, 317, 2009.
How do you do Life Science? http://ki.se
Department of Microbiology, Tumor and Cell Biology – MTC
21
22
Department of Microbiology, Tumor and Cell Biology – MTC
High Impact Publications 2007–2009
Publications with a Journal Impact Factor above 8.
2007
Blood 2007, 109:643–52. Differentiation of
CD1a- and CD1a+ monocyte-derived den–
dritic cells is biased by lipid environment and
PPARg. Gogolak P, Rethi B, Szatmari I, Lanyi
A, Dezso B, Nagy L and Rajnavolgyi E.
Neutrophil
extracellular traps: casting the NET over
pathogenesis. Wartha F, Beiter K, Normark S
and Henriques-Normark B.
Curr Opin Microbiol. 2007, 10:52–6.
Proc Natl Acad Sci USA. 2007, 104:3919–3924.
Filamin B deficiency in mice results in skeletal
malformations and impaired microvascular development. Zhou X, Tian F, Sandzen J,
Cao R, Flaberg E, Szekely L, Cao Y, Ohlsson C,
Bergo MO, Boren J and Akyurek LM.
J Clin Invest. 2007, 117:2362–2368. Angiogenesis
modulates adipogenesis and obesity. Cao Y.
A molecular link
between malaria and Epstein-Barr virus
reactivation. Chêne A, Donati D, GuerreiroCacais AO, Levitsky V, Chen Q, Falk KI, Orem
J, Kironde F, Wahlgren M and Bejarano MT.
PLoS Pathog. 2007, 3:e80.
Proc Natl Acad Sci USA. 2007, 104:12140–5.
Combinatorial protein therapy of angiogenic
and arteriogenic factors remarkably improves
collaterogenesis and cardiac function in pigs.
Lu H, Xu X, Zhang M, Cao R, Bråkenhielm E,
Li C, Lin H, Yao G, Sun H, Qi L,Tang M, Dai
H, Zhang Y, Su R, Bi Y, Zhang Y and Cao Y.
Proc Natl Acad Sci USA. 2007, 104:12907–12.
Clonal success of piliated penicillin nonsusceptible pneumococci. Sjöström K, Blomberg
C, Fernebro J, Dagerhamn J, Morfeldt E, Barocchi MA, Browall S, Moschioni M, Andersson M, Henriques F, Albiger B, Rappuoli R,
Normark S and Henriques-Normark B.
Nat Mater. 2007, 6:673–9. Electronic control of
Ca(2+) signaling in neuronal cells using an
organic electronic ion pump. Isaksson J, Kjäll
P, Nilsson D, Robinson N, Berggren M and
Richter-Dahlfors A.
Adv Cancer Res. 2007, 98:1–16. Why do we not
all die of cancer at an early age? Klein G, Imreh
S and Zabarovsky ER.
Gastroenterology. 2007, 133:808–17. Neuro­
peptide s receptor 1 gene polymorphism
is associated with susceptibility to inflammatory bowel disease. D’Amato M, Bruce
S, Bresso F, Zucchelli M, Ezer S, Pulkkinen
V, Lindgren C, Astegiano M, Rizzetto M,
Gionchetti P, Riegler G, Sostegni R, Daperno
M, D’Alfonso S, Momigliano-Richiardi P,
Torkvist L, Puolakkainen P, Lappalainen
M, Paavola-Sakki P, Halme L, Farkkila M,
Turunen U, Kontula K, Lofberg R, Pettersson
S and Kere J.
Proc Natl Acad Sci USA. 2007, 104:15364–9. IL-20
is an arteriogenic cytokine that remodels
collateral networks and improves functions
of ischemic hind limbs. Tritsaris K, Myren M,
Ditlev SB, Hübschmann MV, van der Blom I,
Hansen AJ, Olsen UB, Cao R, Zhang J, Jia T,
Wahlberg E, Dissing S and Cao Y.
Proc Natl Acad Sci USA. 2007, 104:15835–40.
PfEMP1-DBL1a amino acid motifs in severe
disease states of Plasmodium falciparum
malaria. Normark J, Nilsson D, Ribacke U,
Winter G, Moll K, Wheelock CE, Bayarugaba
J, Kironde F, Egwang TG, Chen Q, Andersson
B and Wahlgren M.
Natural killer cells
trigger differentiation of monocytes into
dendritic cells. Zhang AL, Colmenero P,
Purath U, Teixeira de Matos C, Hueber W,
Klareskog L, Tarner IH, Engleman EG and
Söderström K.
Blood. 2007, 110:2484–93.
J Clin Invest. 2007, 117:2766–2777. Angiogenic
factors FGF2 and PDGF-BB synergistically
promote murine tumor neovascularization
and metastasis. Nissen L J, Cao R, Hedlund
EM, Wang Z, Zhao X, Wetterskog D, Funa K,
Bråkenhielm E and Cao Y.
Gastroenterology. 2007, 133:1499–509. DMBT1
confers mucosal protection in vivo and a deletion variant is associated with crohn’s disease.
Renner M, Bergmann G, Krebs I, End C, Lyer
S, Hilberg F, Helmke B, Gassler N, Autschbach
F, Bikker F, Strobel-Freidekind O, GronertSum S, Benner A, Blaich S, Wittig R, Hudler M,
Ligtenberg AJ, Madsen J, Holmskov U, Annese
V, Latiano A, Schirmacher P, Amerongen AV,
D’Amato M, Kioschis P, Hafner M, Poustka A
and Mollenhauer J.
The world according to MYC. Conference on MYC and the
transcriptional control of proliferation and
oncogenesis. Lüscher B and Larsson LG.
EMBO Rep. 2007, 8:1110–4.
Blood. 2007, 111:2693–703. Leukotriene B4 activates T cells which inhibit B cell proliferation
in EBV infected cord blood derived mononuclear cell cultures. Liu A, Claesson HE,
Mahshid Y, Klein G and Klein E.
2008
J Exp Med. 2008, 205:183–93. Developmental
switch of intestinal antimicrobial peptide
expression. Ménard S, Förster V, Lotz M,
Gütle D, Duerr CU, Gallo RL, HenriquesNormark B, Pütsep K, Andersson M, Glocker
EO and Hornef MW.
Discovery of a
novel class of highly conserved vaccine antigens using genomic scale antigenic fingerprinting of pneumococcus with human
antibodies. Giefing C, Meinke AL, Hanner M,
Henics T, Minh DB, Gelbmann D, Lundberg
U, Senn BM, Schunn M, Habel A, HenriquesNormark B, Örtqvist Å, Kalin M, von Gabain
A and Nagy E.
J Exp Med. 2008, 205:117–31.
Expression of granule-associated proteins in neutrophils from
patients with severe congenital neutropenia.
Andersson M, Karlsson J, Carlsson G and
Pütsep K.
Blood. 2007, 110:2772–3.
Nature. 2007, 449:663. Science & Politics: when
ministers are well primed. Wigzell H.
Department of Microbiology, Tumor and Cell Biology – MTC
23
Citation indicators
Two common bibliometric indicators for MTC publications
2005-2008. Both indicators compare citation counts for
MTC articles to citation counts for world articles from the
same year and in the same research field.
A field normalized citation score average of 1,5 (2005–
2008) means that MTC publications are cited 50% above the
world average. A Top 5% value of 8% (the average of MTC for
2005-2008) means that 8% of MTC publications are among
the worlds 5% most highly cited. Publications from 2009 are
still too new to be evaluated with citation counts.
More information about ­bibliometric indicators used at
­Karolinska Institutet is available at http://ki.se/bibliometrics.
Certain data included herein are derived from the Web
of Science® prepared by THOMSON REUTERS®, Inc.
(Thomson®), Philadelphia, Pennsylvania, USA: ©Copyright
­THOMSON REUTERS® 2010. All rights reserved.
J Exp Med. 2008, 205:63–77. An HIV-1 clade
C DNA prime, NYVAC boost vaccine regimen induces reliable, polyfunctional, and
long-lasting T cell responses. Harari A, Bart
PA, Stöhr W, Tapia G, Garcia M, MedjitnaRais E, Burnet S, Cellerai C, Erlwein O,
Barber T, Moog C, Liljestrom P, Wagner R,
Wolf H, Kraehenbuhl JP, Esteban M, Heeney
J, Frachette MJ, Tartaglia J, McCormack S,
Babiker A, Weber J and Pantaleo G.
Gut. 2008, 57:764–71. Secreted enteric antimicrobial activity localizes to the mucus
surface layer. Meyer-Hoffert U, Hornef
MW, Henriques-Normark B, Axelsson LG,
Midtvedt T, Pütsep K and Andersson M.
The challenges of eliciting neutralizing antibodies
to HIV-1 and to influenza virus. Karlsson
Hedestam GB, Fouchier RA, Phogat S, Burton
DR, Sodroski J and Wyatt RT.
Nat Rev Microbiol. 2008, 6:143–55.
2,2
Field Normalized Citation Score
rococcus faecalis from newborn babies regulate endogenous PPARg activity and IL-10
levels in colonic epithelial cells. Are A, Aronsson L, Wang S, Greicius G, Lee YK, Gustafsson
JA, Pettersson S and Arulampalam V.
Nat Cell Biol. 2008, 10:211–9. Membrane nano-
tubes physically connect T cells over long
distances presenting a novel route for HIV-1
transmission. Sowinski S, Jolly C, Berninghausen O, Purbhoo MA, Chauveau A,
Köhler K, Oddos S, Eissmann P, Brodsky FM,
Hopkins C, Onfelt B, Sattentau Q and Davis
DM.
24
Department of Microbiology, Tumor and Cell Biology – MTC
Top 5%
11%
2,0
10%
1,8
9%
1,6
8%
1,4
7%
1,2
6%
1,0
5%
0,8
4%
0,6
3%
0,4
2%
0,2
1%
0%
0,0
2005
2006
Genome Res. 2008, 18:370–9. Segmental duplications and evolutionary plasticity at tumor
chromosome break- prone regions. DaraiRamqvist E, Sandlund A, Müller S, Klein G,
Imreh S and Kost-Alimova M.
J Clin Invest. 2008, 118:913–23. VEGF-B inhibits apoptosis via VEGFR-1-mediated suppression of the expression of BH3-only protein
genes in mice and rats. Li Y, Zhang F, Nagai
N, Tang Z, Zhang S, Scotney P, Lennartsson J, Zhu C, Qu Y, Fang C, Hua J, Matsuo O,
Fong GH, Ding H, Cao Y, Becker KG, Nash A,
Heldin CH and Li X.
Proc Natl Acad Sci USA. 2008, 105:4358–63.
Helicobacter pylori evolution during progression from chronic atrophic gastritis to gastric
cancer and its impact on gastric stem cells.
Giannakis M, Chen SL, Karam SM, Engstrand
L and Gordon JI.
Proc Natl Acad Sci USA. 2008, 105:5489–94. EBV-
Proc Natl Acad Sci USA. 2008, 105:1943–8. Ente-
World Average
encoded EBNA-6 binds and targets MRS18–2
to the nucleus, resulting in the disruption of
pRb-E2F1 complexes. Kashuba E, Yurchenko
M, Yenamandra SP, Snopok B, Isaguliants M,
Szekely L and Klein G.
Nat Immunol. 2008, 9, 477–80. Natural killer cell
recognition of missing self. Kärre K.
Twin ushers guide pili
across the bacterial outer membrane. Daniels
R and Normark S.
Cell. 2008, 133:574–6.
2007
2008
Blood. 2008, 112:1461–71. Neutrophil secretion products pave the way for inflammatory
monocytes. Soehnlein O, Zernecke A, Eriksson EE, Rothfuchs AG, Pham CT, Herwald H,
Bidzhekov K, Rottenberg ME, Weber C and
Lindbom L.
Priming of T cells
to Fas-mediated proliferative signals by interleukin-7. Rethi B, Vivar N, Sammicheli S,
Fluur C, Ruffin N, Atlas A, Rajnavolgyi E and
Chiodi F.
Blood. 2008, 112:1195–204.
Adv Cancer Res. 2008, 100:113–131. Molecular
mechanisms and therapeutic development of
angiogenesis inhibitors. Cao Y.
Proc Natl Acad Sci USA. 2008, 105:10167–72.
FOXC2 controls Ang-2 expression and
modulates angiogenesis, vascular patterning,
remodeling, and functions in adipose tissue.
Xue Y, Cao R, Nilsson D, Chen S, Westergren
R, Hedlund EM, Martijn C, Rondahl L, Krauli
P, Walum E, Enerbäck S and Cao Y.
Proc Natl Acad Sci USA. 2008, 105:13203–5.
A review of Judah Folkman’s remarkable
achievements in biomedicine. Cao Y and
Langer R.
Proc Natl Acad Sci USA. 2008, 105:15028–33.
Inflammation and autoimmunity caused by a
SHP1 mutation depend on IL-1, MyD88, and
a microbial trigger. Croker BA, Lawson BR,
Berger M, Eidenschenk C, Blasius AL, Moresco
EM, Sovath S, Cengia L, Shultz LD, Theofilopoulos AN, Pettersson S and Beutler BA.
PLoS Pathog. 2008, 4:e1000171. B cell recognition on the conserved HIV-1 co-receptor
binding site is altered by endogenous primate
CD4. Forsell MN, Dey B, Mörner A, Svehla K,
O’dell S, Högerkorp CM, Voss G, Thorstensson R, Shaw GM, Mascola JR, Karlsson
Hedestam GB, Wyatt RT. Högerkorp CM,
Voss G, Thorstensson R, Shaw GM, Mascola
JR, Karlsson Hedestam GB and Wyatt RT.
2009
Cell Metab. 2009, 9:99–109. Hypoxia-independ-
ent angiogenesis in adipose tissues during cold
acclimation. Xue Y, Petrovic N, Cao R, Larsson
O, Lim S, Chen S, Feldmann HM, Liang Z, Zhu
Z, Nedergaard J, Cannon B and Cao Y.
Proc Natl Acad Sci USA. 2009, 106:7939–44.
Timing of HAART defines the integrity of
memory B cells and the longevity of humoral
responses in HIV-1 vertically-infected children. Pensieroso S, Cagigi A, Palma P, Nilsson
A, Capponi C, Freda E, Bernardi S, Thorstensson R, Chiodi F and Rossi P.
Proc Natl Acad Sci USA. 2009, 106:859-63
Towards a genetics of cancer resistance. Klein G.
Mycobacterial glycoconjugates as vaccine candidates
against tuberculosis. Källenius G, Pawlowski
A, Hamasur B and Svenson SB.
Trends Microbiol. 2008, 16:456–62.
Nat Rev Microbiol. 2008, 6:827–37. The rise and
fall of bacterial clones: Streptococcus pneumoniae. Henriques-Normark B, Blom­berg C,
Dagerhamn J, Bättig P and Normark S.
Circ Res. 2008, 103:1092–9. Vascular endothelial
growth factor-A and platelet-derived growth
factor-B combination gene therapy prolongs
angiogenic effects via recruitment of interstitial mononuclear cells and paracrine effects
rather than improved pericyte coverage of
angiogenic vessels. Korpisalo P, Karvinen H,
Rissanen TT, Kilpijoki J, Marjomäki V, Baluk
P, McDonald DM, Cao Y, Eriksson U, Alitalo K
and Ylä-Herttuala S.
Cytokine Growth Factor Rev. 2008, 19:395–404.
Interferon and cytokine responses to Crimean
Congo hemorrhagic fever virus; an emerging
and neglected viral zonoosis. Weber F and
Mirazimi A.
Proc Natl Acad Sci USA. 2008, 105:18513–8.
Anti-VEGF agents confer survival advantages
to tumor-bearing mice by improving cancerassociated systemic syndrome. Xue Y, Religa P,
Cao R, Hansen AJ, Lucchini F, Jones B, Wu Y,
Zhu Z, Pytowski B, Liang Y, Zhong W, Vezzoni
P, Rozell B and Cao Y.
Blood. 2008, 112:4401–10. Altered expression
of the receptor-ligand pair CXCR5/CXCL13
in B-cells during chronic HIV-1 infection.
Cagigi A, Mowafi F, Phuong Dang LV, TennerRacz K, Atlas A, Grutzmeier S, Racz P, Chiodi
F and Nilsson A.
Great times for small
molecules: c-AMP, a second messenger candidate in Bacteria and Archaea. Römling U.
Sci Signal. 2008, 1:pe39.
Positive and negative
modulation of angiogenesis by VEGFR1
ligands. Cao Y.
Sci Signal. 2009, 2:re1.
Blood. 2009, 113:2434–41. The strength of
inhibitory input during education quantitatively tunes the functional responsiveness
of individual natural killer cells. Brodin P,
Lakshmikanth T, Johansson S, Kärre K and
Höglund P.
Nat Rev Genet. 2009, 10:269–76. Replication
timing and epigenetic reprogramming of
gene expression: a two-way relationship?
Göndör A and Ohlsson R.
Proc Natl Acad Sci USA. 2009, 106:6152–7.
VEGF-B is dispensable for blood vessel growth
but critical for their survival, and VEGF-B
targeting inhibits pathological angiogenesis.
Zhang F, Tang Z, Hou X, Lennartsson J, Li Y,
Koch AW, Scotney P, Lee C, Arjunan P, Dong L,
Kumar A, Rissanen TT, Wang B, Nagai N, Fons
P, Fariss R, Zhang Y, Wawrousek E, Tansey G,
Raber J, Fong GH, Ding H, Greenberg DA,
Becker KG, Herbert JM, Nash A, Yla-Herttuala S, Cao Y, Watts RJ and Li X.
NCRs and
DNAM-1 mediate NK cell recognition and
lysis of human and mouse melanoma cell
lines in vitro and in vivo. Lakshmikanth T,
Burke S, Ali TH, Kimpfler S, Ursini F, Ruggeri
L, Capanni M, Umansky V, Paschen A, Sucker
A, Pende D, Groh V, Biassoni R, Höglund P,
Kato M, Shibuya K, Schadendorf D, Anichini
A, Ferrone S, Velardi A, Kärre K, Shibuya A,
Carbone E and Colucci F.
Cancer Cell. 2009, 15:171–83. MDM2-Dependent downregulation of p21 and hnRNP K
provides a switch between apoptosis and
growth arrest induced by pharmacologically
activated p53. Enge M, Bao W, Hedström E,
Jackson SP, Moumen A and Selivanova G.
Proc Natl Acad Sci USA. 2009, 106:11966–71. The
proapoptotic function of SAP provides a clue
to the clinical picture of X-linked lymphoproliferative disease. Nagy N, Matskova L, Kis LL,
Hellman U, Klein G and Klein E.
J Clin Invest. 2009, 119:1251–63.
Ablation of key
oncogenic pathways by RITA-reactivated p53
is required for efficient apoptosis. Grinkevich VV, Nikulenkov F, Shi Y, Enge M, Bao W,
Maljukova A, Gluch A, Kel A, Sangfelt O and
Selivanova G.
Cancer Cell. 2009, 15:441–53.
Proc Natl Acad Sci USA. 2009, 106:14478–83.
Chromosome 14 transfer and functional
studies identify a candidate tumor suppressor gene, mirror image polydactyly 1, in
nasopharyngeal carcinoma. Cheung AK,
Lung HL, Ko JM, Cheng Y, Stanbridge EJ,
Zabarovsky ER, Nicholls JM, Chua D, Tsao
SW, Guan XY and Lung ML.
PLoS Pathog. 2009, 5:e1000584. Getting to grips
with strangles: an effective multi-component
recombinant vaccine for the protection of
horses from Streptococcus equi infection. Guss
B, Flock M, Frykberg L, Waller AS, Robinson
C, Smith KC and Flock JI.
Nature. 2009, 461:212–7. Chromosome crosstalk in three dimensions. Göndör A and Ohlsson R.
Department of Microbiology, Tumor and Cell Biology – MTC
25
Proc Natl Acad Sci USA. 2009, 106:17505–10.
Malignant cell-derived PlGF promotes
normalization and remodeling of the tumor
vasculature. Hedlund EM, Hosaka K, Zhong
Z, Cao R and Cao Y.
Semin Cancer Biol. 2009, 19:338–43. Improvement of antiangiogenic cancer therapy by
understanding the mechanisms of angiogenic
factor interplay and drug resistance. Cao Y,
Zhong W and Sun Y.
Semin Cancer Biol. 2009, 19:277–8.
Angiogen-
esis in malignancy. Cao Y.
Cell. 2009, 139:679–92. Dissociation of EphB2
signaling pathways mediating progenitor
cell proliferation and tumor suppression.
Genander M, Halford MM, Xu NJ, Eriksson M, Yu Z, Qiu Z, Martling A, Greicius G,
Thakar S, Catchpole T, Chumley MJ, Zdunek
S, Wang C, Holm T, Goff SP, Pettersson S,
Pestell RG, Henkemeyer M and Frisén J.
Proc Natl Acad Sci USA. 2009, 106:19866–71.
MRPS18–2 protein immortalizes primary rat
embryonic fibroblasts and endows them with
stem cell-like properties. Kashuba E, Pavan
Yenamandra S, Deoram Darekar S, Yurchenko M, Kashuba V, Klein G and Szekely L.
Proc Natl Acad Sci USA. 2009 106:18408–13.
Nitric oxide permits hypoxia-induced
lymphatic perfusion by controlling arteriallymphatic conduits in zebrafish and glass
catfish. Dahl Ejby Jensen L, Cao R, Hedlund
EM, Söll I, Lundberg JO, Hauptmann G, Steffensen JF and Cao Y.
Genes Dev. 2009, 23:2598–603. Nonallelic trans-
vection of multiple imprinted loci is organized by the H19 imprinting control region
during germline development. Sandhu KS,
Shi C, Sjölinder M, Zhao Z, Göndör A, Liu L,
Tiwari VK, Guibert S, Emilsson L, Imreh MP
and Ohlsson R.
Proc Natl Acad Sci USA. 2009, 106:19485–90.
Hypoxia-induced pathological angiogenesis
mediates tumor cell dissemination, invasion,
and metastasis in a zebrafish tumor model.
Lee SL, Rouhi P, Dahl Jensen L, Zhang D, Ji H,
Hauptmann G, Ingham P and Cao Y.
Semin Cancer Biol. 2009, 19:411–20. Endemic
Burkitt’s lymphoma as a polymicrobial disease: new insights on the interaction between Plasmodium falciparum and
Epstein-Barr virus. Chêne A, Donati D, Orem
J, Mbidde ER, Kironde F, Wahlgren M and
Bejarano MT.
Semin Cancer Biol. 2009 19:407–10. To the
genesis of Burkitt lymphoma: regulation of
apoptosis by EBNA-1 and SAP may determine the fate of Ig-myc translocation carrying
B lymphocytes. Nagy N, Klein G and Klein E.
Connections between MTC research groups and their main research areas (as identified with MeSH terms). The MeSH headings must occur over 10 times
in MTC articles to be shown and the connection to each individual research group must be at least 2 articles strong to show up as a line. The MeSH Headings
used in this analysis were from MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine. NLM represents that its data were formulated with a
reasonable standard of care. Except for this representation, NLM makes no representation or warranties, expressed or implied. This includes, but is not limited
to, any implied warranty of merchantability or fitness for a particular purpose, with respect to the NLM data, and NLM specifically disclaims any such warranties
and representations. Certain data included herein are derived from the Web of Science ® prepared by THOMSON REUTERS ®, Inc. (Thomson®), Philadelphia,
Pennsylvania, USA: ©Copyright THOMSON REUTERS® 2010. All rights reserved.
26
Department of Microbiology, Tumor and Cell Biology – MTC
Schistosome worm sections “Andyfied” by Ewert Linder.
Department of Microbiology, Tumor and Cell Biology – MTC
27
Research Reports Immunobiology
Immunology Research
MTC is one of the most active Departments at the Karolinska Institutet (KI) in
the field of immunology. This is in no small
part thanks to our excellent animal facility,
which harbors a number of animal models
for human disease and an extensive panel
of genetically modified mice of interest to
immunologists. Many mouse models to study
immune responses to infectious agents as
well as anti-tumor responses and autoimmunity are available at MTC and the number of
mouse strains housed at the facility is continuously expanding.
While the majority of the immunobiology
projects at MTC rely on pre-clinical experimental systems, there is also a focus on translational research. Several investigators have a
clinical background and/or collaborate with
clinicians at Karolinska University Hospital or the Stockholm South General Hospital (Södersjukhuset), which ensures access
to important expertise and unique patient
samples. Also of note, the KI-wide research
network “KI immunology and inflammation (KiiM)” was initiated from MTC and
researchers from MTC play active roles in
managing this network. The regular KiiM
Gunnel Biberfeld
Francesca Chiodi
retreat is an increasingly popular event, which
has attracted a number of distinguished international speakers over the years.
Natural killer cell immunobiology
A highly visible research area at MTC is the
field of natural killer (NK) cell immunobiol-
Petter Höglund
Mikael Jondal
Gunilla Karlsson
Hedestam
ogy. Several research groups study NK cells
to understand their development and function, as well as the mechanisms underlying
the process of NK cell education. The activities of NK cells are regulated by a number
of activating and inhibitory receptors that
can be manipulated to alter NK cell func-
Project leaders:
Louise Berg
Maria Johansson
Gerald McInerney
Björn Önfelt
Bence Rethi
Markus Sköld
Jonas Sundbäck
Klas Kärre
28
Peter Liljeström
Markus Maeurer
Department of Microbiology, Tumor and Cell Biology – MTC
Martin Rottenberg
Hans Wigzell
Research Reports Immunobiology
Photo: Lasse Skog
tion. Groups at MTC have shown that by
blocking inhibitory receptors expressed by
NK cell, enhanced rejection of leukemia
cells or enhanced anti-viral activity against
HIV-1 or herpes viruses infected cells may
be achieved. NK cells in organ-specific
autoimmunity are also studied, such as NK
cells in the pancreas in a mouse model of
human type I diabetes. Furthermore, new
technological approaches for understanding how NK cells and T cells communicate
with target cells to exert their function were
recently established at MTC.
Human immune responses
Many additional basic immunology questions are also investigated at MTC, such as
how immunological aging occurs and how
this relates to the loss of thymic tissue. Efforts
are also underway to extend the mouse model
beyond its current use by engrafting human
CD34+ hematopoietic stem cells to develop
mice harboring human immune cells to study
human immune responses to pathogens such
as mycobacteria.
A Mycobacterium tuberculosis (Mtb)
aerosol infection model has also been set up,
(HIV-1). The mechanisms through which T
which mimics the natural route of infection
and B cells are damaged during HIV-1 infecallowing the analysis of monocyte differentiation are studied and amongst other findings it
tion and function during
was shown that activationMtb infection. A further
induced apoptosis increase
“While the majority of the
effort in the field of Mtb is
with disease progression.
immunobiology projects
to identify target antigens
The need to halt the
at MTC rely on pre-clinical
within the Mtb proteome
global spread of HIV-1 calls
experimental systems,
for use in diagnostics
for improved approaches to
there is also a focus on
or as vaccine antigens.
induce protective immune
translational research.”
Additional bacteria and
responses against this
parasites that are studgenetically diverse virus.
ied at MTC include the bacteria Salmonella,
This goal is approached by several groups and
Chlamydia, Listeria and Pneumococci and
has resulted in the development of recomthe parasites Trypanozomes and Leishmania,
binant viral vaccine vectors and engineered
all of which have evolved means to interfere
protein antigens, some of which have entered
with the host immune response.
clinical trials already. The need to stimulate
responses to highly conserved and often subdominant viral epitopes puts special demands
The mechanisms of HIV-1 infection
on the design of vaccine immunogens and
The interaction between viruses and the host
on understanding how to focus T and B cell
immune system is another main area within
responses on relevant antigenic determinants.
the field of immunology at MTC. An imporThis is an active area of investigation at MTC,
tant viral pathogen that causes chronic infecwhich also covers the mechanism of action of
tion and, in the absence of anti-retroviral thervaccine adjuvants.
apy, renders the host immune system defective is human immunodeficiency virus type 1
Gunilla Karlsson Hedestam
Department of Microbiology, Tumor and Cell Biology – MTC
29
Research Reports Immunobiology
Translational studies of
NK cell function in HIV and
herpes virus infection
We investigate natural killer (NK)
cell function in patients with HIV
infection and in healthy children
in relation to early encounter of
herpes virus infections. NK cells
Louise Berg
are innate lymphocytes regulated
[email protected]
by expression of a multitude of
inhibitory and activating receptors, interacting with endogenous or virally encoded cell surface
bound ligands.
Although antiretroviral therapy greatly delays disease progression
in HIV, there is still no cure for infection. We focus on investigating
NK cell based treatment in HIV. Susanne Johansson has characterised the function and receptor expression of NK cells from viremic
patients, patients under antiretroviral treatment and patients who
efficiently control viremia without treatment. She has seen that blockade of NK cell inhibitory receptors increases NK cell recognition of
HIV infected autologous T cells in vitro. We are presently studying
antibody dependent cellular cytotoxicity (ADCC) in HIV, and aim to
investigate whether inhibitory receptor blockade in combination with
ADCC may be an effective treatment strategy in HIV. This project is
run in collaboration with the Stockholm South General Hospital.
NK cells have been
extensively studied in
herpes virus infection,
mainly with cytomegalovirus (CMV) (Figure).
In a collaboration with
researchers at Stockholm University, we
have investigated NK
cells during childhood
Immune evasion strategies of
and seen that recepcytomegalovirus.
tor expression develops
during the first years of life. In addition, we found that children who
encounter herpes virus infections early in life have a decreased monocyte induced NK cell function, as seen during asymptomatic chronic
EBV and CMV infection. This may bear impact on the development
of allergy, which is less frequent in children encountering early infections with herpes viruses. We conclude that environmental factors,
such as chronic herpes virus infections, shape innate immune functions in early life. This project is run in collaboration with clinicians
from the Department of Pediatrics, Sachs’ Children’s Hospital at the
Stockholm South General Hospital.
Prevention of mother-tochild transmission of HIV
in Dar es Salaam, Tanzania
My research group has collaborated
with the Muhimbili University of
Health and Allied Sciences in Dar es
Salaam, Tanzania for more than 20
years in the Swedish International
Gunnel Biberfeld
Development Cooperation Agency
[email protected]
(Sida)-supported TANSWED HIV
program. Two of the main projects
in this program are: “Prevention of mother-to-child transmission of
HIV” and “Evaluation of HIV vaccines”.
About 400,000 children become HIV-infected each year in subSaharan Africa, the majority through mother-to-child transmission
(MTCT) of HIV. Prophylactic antiretroviral (ARV) treatment is given
to HIV-infected mothers and their infants before and around delivery to reduce the risk of early MTCT of HIV in developing countries.
However, this prophylactic treatment does not prevent postnatal HIV
transmission through breast milk. Replacement feeding from birth is
not feasible, safe or affordable in most resource-limited settings. We
have performed two studies in Dar es Salaam to evaluate prevention
of breast milk transmission of HIV
by giving extended ARV prophylaxis
to HIV-infected mothers or to their
infants during breast feeding for 6
months after delivery. ARV treatment
continued for mothers who needed
treatment for their own health.
These studies showed that infant
or maternal ARV prophylaxis for 6
months during breast feeding resulted
in a similar low risk of acquisition of
infant HIV infection between 6 weeks
A mother and her child waitand 6 months (1%) and a similar low
ing for examination in the
research clinic in Dar es
cumulative infant HIV infection rate at
Salaam.
6 months of age (5%). Based on these
and other similar studies in sub-Saharan Africa, the World Health
Organization has issued new guidelines for prevention of postnatal
infection in infants of HIV-infected mothers who do need treatment
for their own health recommending ARV prophylaxis to either the
mother or the infant during the whole duration of breast feeding.
Selected publications
l
Selected publications
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30
Sundström Y, Nilsson C, Lilja G, Kärre K, Troye-Blomberg M and
Berg L. The expression of human NK cell receptors in early life.
Scand J Immunol 66, 335, 2007.
Shagafian-Hedengren S, Sundström Y, Sohlberg E, Nilsson C,
Linde A, Troye-Blomberg M, Sverremark-Ekström E and Berg L.
Herpesvirus seropositivity in childhood associates with decreased
monocyte-induced NK-cell IFN-g production. J Immunol, 182,
2511, 2009.
Johansson S, Hinkula J, Hejdeman B, Johansson M, Wahren B,
Wagtmann N R, Kärre K and Berg L. NK-cell activation by KIR
binding antibody 1-7F9 and response to in vitro HIV infected
autologous cells in viremic and controller HIV infected patients.
Clin Immunol 134, 158, 2010.
Department of Microbiology, Tumor and Cell Biology – MTC
l
l
Kilewo C, Karlsson K, Massawe A, Lyamuya E, Swai A, Mhalu F,
Biberfeld G and the Mitra Study Team. Prevention of mother-tochild transmission of HIV-1 through breast-feeding by treating
infants prophylactically with lamivudine in Dar es Salaam, Tanzania: the Mitra Study. J Acquir Immune Defic Syndr 48, 315, 2008.
Sandström E, Nilsson C, Hejdeman B, Bråve A, Bratt G, Robb
M, Cox J, Vancott T, Marovich M, Stout R, Aboud S, Bakari M,
Pallangyo K, Ljungberg K, Moss B, Earl P, Michael N, Birx D, Mhalu
F, Wahren B, Biberfeld G and the HIV Immunogenicity Study 01/02
Team. Broad immunogenicity of a multigene, multiclade HIV-1
DNA vaccine boosted with heterologous HIV-1 recombinant
modified vaccinia virus Ankara. J Infect Dis 198, 1482, 2008.
Kilewo C, Karlsson K, Ngarina M, Massawe A, Lyamuya E, Swai A,
Lipyoga R, Mhalu F, Biberfeld G and the Mitra Plus Study Team.
Prevention of mother-to-child transmission of HIV-1 through
breastfeeding by treating mothers with triple antiretroviral therapy in Dar es Salaam, Tanzania: the Mitra Plus study. J Acquir
Immune Defic Syndr 52, 406, 2009.
Research Reports Immunobiology
Understanding the mechanism
of damage induced by HIV-1 to
T and B cells may lead to faster
development of a HIV-1 vaccine
Only a proportion of T
cells which die during
HIV-1 infection are directly
infected by the virus; and
memory B cells, whose
number is reduced during
Francesca Chiodi
HIV-1 infection, are not a
[email protected]
direct target for HIV-1. My
group aims to understand
the mechanism(s) through which T and B cells are damaged during
HIV-1 infection. To determine whether dysfunctional cell homing
may play a role in impairments of B cell responses, the expression of
chemokine receptors and their respective ligands were examined on
B cells. We reported a decreased CXCR5 expression and an increased
expression of CXC chemokine ligand 13 (CXCL13) on B cells from
HIV-1 infected patients, in association with low CD4 T cell counts.
Altered expression of the chemokine receptor-ligand pair, CXCR5/
CXCL13, may thus participate in the establishment of B cell dysfunctions during chronic HIV-1 infection.
We reasoned that an early control of viral replication through
anti-viral treatment could preserve the normal development of the
memory B cell compartment and responses to routine childhood
vaccines. Accordingly, we evaluated the effects of different medication schedules in 70 HIV-1 vertically-infected pediatric subjects on B
cell functions. We showed that timing of medication initiation is the
major factor predicting the longevity of B cell responses in vaccinated
HIV-1-infected children.
The Fas receptor,
primarily associated with
T and B cell depletion in
lymphopenic
settings,
was previously shown to
contribute to compensatory Tcell expansion. As
IL-7 levels are elevated
during HIV-1 infection in association with
increased Fas expression,
we analyzed whether IL-7
influences Fas-mediated proliferative signals in T cells. We showed
that IL-7 is able to increase the efficacy of Fas to induce proliferation
of suboptimally activated T cells. High IL-7 levels may simultaneously
induce sensitivity to Fas-mediated apoptosis in non-activated T cells
and increase Fas-induced co-stimulatory signals in T cells recognizing
low-affinity antigens.
Dissecting natural killer
cell tolerance in models
of transplantation and
autoimmunity
Natural killer (NK) cells mediate graft-versus-leukemia effects
after stem cell transplantation
and contribute to autoimmunity.
NK cell functions are regulated
by MHC class I molecules, which
Petter Höglund
also control development of NK
[email protected]
cells, a process we have termed
“NK cell education”. A major
objective of our work is to identify the molecular mechanisms for this
education process using murine model systems.
The inhibitory receptors, called Ly49 in mice, and their MHC
ligands are both encoded in multigenic gene families and display
polymorphisms and crossreactivities. By studying transgenic mice
expressing individual MHC class I alleles in isolation, pairs and
triplets using polychromatic flow cytometry protocols, we are able to
dissect the system despite this complexity. We have found that some
MHC class I alleles have a strong “educating” impact on the NK cell
system, while some alleles have a weak impact. In addition, we have
shown that NK cell education is regulated quantitatively in individual
NK cells. Several novel insights into the process of NK cell education
have been delinated, which are further studied in clinical collaborations involving patients undergoing stem cell transplantation against
hematological malignancy.
In a second objective, NK cells in organ-specific autoimmunity is
studied. Non-obese diabetic (NOD) mice develop a type I diabetes
similar to the human disase.
We focus on NK cells in the
target organ, the pancreas.
Pancreatic NK cells show an
altered phenotype compared
to NK cells in the hematopoietic system and localize both
in the islets of Langerhans
and in the exocrine part of
the organ. The fraction of NK
cells inside the islets increased
as disease progresses, implyAn islet of Langerhans with infiltrating
ing a local role of NK cells in
NK cells. Red = Insulin; blue = T cells
and green = NK cells.
diabetes pathogenesis. NK cell
depletion studies suggest that
NK cells indeed promote the development of diabetes, and our future
work focusses on identifying the cellular and molecular regulation of
this effect.
Selected publications
Selected publications
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l
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Cagigi A, Mowafi F, Phuong Dang LV, Tenner-Racz K, Atlas A,
Grutzmeier S, Racz P, Chiodi F and Nilsson A. Altered expression of the receptor-ligand pair CXCR5/CXCL13 in B cells during
chronic HIV-1 infection. Blood 112, 4401, 2008.
Rethi B, Vivar N, Sammicheli S, Fluur C, Ruffin N, Atlas A, Rajnavolgyi E and Chiodi F. Priming of T cells to Fas-mediated proliferative signals by interleukin-7. Blood 15, 112, 2008.
Pensieroso S, Cagigi A, Palma P, Nilsson A, Capponi C, Freda
E, Bernardi S, Thorstensson R, Chiodi F and Rossi P. Timing of
HAART defines the integrity of memory B cells and the longevity
of humoral responses in HIV-1 vertically-infected children. Proc
Natl Acad Sci USA 106, 7939, 2009.
l
l
l
Johansson S, Salmon-Divon M, Johansson MH, Pickman Y,
Brodin P, Kärre K, Mehr R and Höglund P. Probing natural killer
cell education by Ly49 receptor expression analysis and computational modelling in single MHC class I mice. PLoS One 4, e6046,
2009.
Brodin P, Lakshmikanth T, Johansson S, Kärre K and Höglund P.
The strength of inhibitory input during education quantitatively
tunes the functional responsiveness of individual natural killer
cells. Blood 113, 2434, 2009.
Brauner H, Elemans M, Lemos S, Broberger C, Holmberg D, Flodström-Tullberg, M, Kärre K and Höglund P. Distinct Phenotype
and function of natural killer cells in the pancreas of non obese
diabetic mice. J Immunol 184, 2272, 2010.
Department of Microbiology, Tumor and Cell Biology – MTC
31
Research Reports Immunobiology
Natural killer cells – anti-tumor
reactivity and self-tolerance
during inhibitory receptor
blockade
Our work aims at developing and using mouse
models where antileukemic
reactivity can be improved
by employing natural killer
(NK) cells. NK cells are
Maria Johansson
lymphocytes in the innate
[email protected]
immune system and their
activity is regulated by activating and inhibitory receptors. We study well characterized inhibitory receptors for self MHC class I on NK cells. By blocking these
receptors, one can induce NK cell rejection of leukemia cells in mice.
We found that this anti-tumor reactivity can be potentiated by exogenous IL-2. In addition, we investigated the risk of breaking NK cell
tolerance to normal cells by such treatment, and found no signs of
autoreactivity (Vahlne G et al., 2008). The presence of NK cell activating ligands on normal hematopoietic cells suggest that also such cells
should be killed during
inhibitory receptor blockade. We are now further
investigating the basis for
the robust self-tolerance
during treatment. We are
also developing mouse
models for utilization of
NK cells in hematopoietic
stem cell (HSC) transplantation (see report by
Blocking of inhibitory receptors on NK cells
Klas Kärre). In closely
induce killing of tumor cells
related projects we are
investigating if blockade
of inhibitory NK cell receptors could induce beneficial reactivity
against virus infected cells by mouse as well as by human NK cells (see
report by Louise Berg).
Sensitive and accurate assays are a cornerstone of experimental research. We are developing single cell assays for cellular effector
functions. We have adapted to the mouse NK cell system, a method
for measuring exocytosis of secretory lysosomes (degranulation)
utilizing cell surface detection of the marker CD107a (Vahlne G et al.,
2008). By the use of multicolor flow cytometry, this analysis can be
combined with analysis of other functions such as cytokine production as well as with phenotypic analysis.
The involution of
the thymus gland
Immunological aging occurs at many levels
within the immune system but one decisive
factor is the loss of thymic tissue (thymic
involution) as thymus is the only site where
Mikael Jondal new T cells can be formed. The decline in
[email protected]
the formation of T cells leads to an agerelated loss of function within the cellular
immune system. Thymic involution is especially related to the formation of sex hormones, formed during puberty.
In mice, we have found that both epithelial and lymphoid (thymocytes) cells in the thymus can synthesize corticosterone (CS), a glucocorticoid which can kill lymphoid cells by the initiation of a cellular
suicide program called “apoptosis”. For this reason we are testing the
hypothesis that an age-related CS synthesis in the thymus contribute
to thymic involution.
We have found that CS synthesis in pre-puberty mice occurs mainly
in epithelial cells whereas at puberty, thymocytes start to synthesize
CS. This synthesis can be triggered by the male sex hormone testosterone in mice that have no endogenous production of this hormone
(castrated mice). In addition, the pharmacological inhibition of local
CS synthesis in thymocytes in old mice resulted in a strong regeneration of thymic tissue (see Figure) resulting in an increased formation
of thymocytes. We are presently investigating how the reversal of
thymic involution impacts the diversity and function of T cells in the
peripheral immune system.
As the immune system declines with age, infectious disease is a
common cause of death in the elderly. This is becoming an issue of
growing medical importance as the mean life expectancy is rapidly
increasing, from around 40 to 80 years during the last 200 years. We
believe that regeneration of thymic tissue in the elderly may boost
cellular immunity and thus contribute to a longer and healthier life
in this population. The inhibition of local glucocorticoid synthesis in
the thymus may be one way to achieve this.
Selected publications
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32
Vahlne G, Becker S, Brodin P and Johansson M.H. IFN-g production and degranulation are differentially regulated in response to
stimulation in murine NK cells. Scand J Immunol 67,1, 2008.
Johansson, S, Salmon-Divon M, Johansson M H, Pickman Y,
Brodin P, Kärre K, Mehr R and Höglund P. Probing natural killer
cell education by Ly49 receptor expression analysis and computational modeling in single MHC class I mice. PLoS ONE 4, e6046,
2009.
Johansson S.E, Hejdeman B, Hinkula J, Johansson M.H, Romagné
F, Wahren B, Wagtmann N R, Kärre K and Berg L. NK cell activation by KIR-binding antibody 1-7F9 and response to HIV-infected
autologous cells in viremic and controller HIV-infected patients.
Clin Immunol 134, 158, 2010.
Department of Microbiology, Tumor and Cell Biology – MTC
The effect of inhibition of the local glucocorticoid synthesis in the thymus. In
treated mice there is a pronounced increase in thymocyte numbers seen as
an expansion of the cortical area of the gland.
Selected publications
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Qiao S, Chen L, Okret S and Jondal M. Age-related synthesis of
glucocorticoids in thymocytes. Exp Cell Res 314, 3027, 2008.
Chen L, Fredholm BB and Jondal M. Adenosine, through the A1
receptor, inhibits vesicular MHC class I cross-presentation by resting DC. Mol Immunol 45, 2247, 2008.
Qiao S, Okret S and Jondal M. Thymocyte-synthesized glucocorticoids play a role in thymocyte homeostasis and are down-regulated by adrenocorticotropic hormone. Endocrinology 150, 4163,
2009.
Research Reports Immunobiology
Virus-cell interactions and
induction of anti-viral B cell
responses
MHC class I molecules: recognition
by NK cells and T cells, novel
functions in nerve cells
We use experimental systems
to investigate how immune
responses are induced following
virus infection and immunization. The quality of adaptive
Gunilla Karlsson Hedestam
immune responses is determined
[email protected]
by early events following the
initial encounter between the
pathogen and the host. We believe that these processes can be manipulated at multiple levels to improve the design of human vaccines. We
are particularly interested in elicitation of anti-viral B cell responses
as neutralizing antibodies are the correlates of protection for most
vaccines that provide protection against infection and disease. Several
projects in the group concern antibody responses against the HIV-1
envelope glycoproteins (Env), for which we have created a range of
recombinant variants that lack or do not lack defined structural determinants. We recently showed that these immunogens stimulate B cell
responses that are markedly different, both in terms of the specificities and the functions of the elicited antibodies. By further developing these approaches we hope to focus vaccine-induced responses
on relevant neutralizing antibody targets to enhance the protective
effect. To facilitate these analyses, we have developed new methods
to enumerate different classes of B cells in immunized and infected
subjects. In a different project, we investigate the intrinsic adjuvant
effect of viruses, primarily using Semliki Forest virus (SFV), but also
other RNA viruses. In one approach, we use SFV for an in vivo screen in
mice to search for non-redundant cellular pathways that are required
for elicitation of B cell responses against virus-encoded antigens. We
have identified a number of
candidate genes that are now
under further investigation.
While many projects in the
group are based on in vivo
models, we also study direct
interactions between viruses
and host cells in vitro to
understand anti-viral signaling events, stress responses
and antigen presentation, all
of which influence the eliciThe HIV-1 envelope core structure with
tation of adaptive immune
key amino acid residues highlighted.
responses.
Natural killer (NK) cells
express a panel of germ
line encoded activating
and inhibitory receptors.
Most inhibitory ligands
Klas Kärre are expressed by normal
[email protected]
cells, e.g. MHC class I
molecules. Some activating ligands are also expressed on normal cells, while others are
expressed only in stressed, transformed or infected cells. NK cells
make decisions by integrating the input via these receptors. The
consequence when activation prevails is release of cytokines (e.g.
IFN-γ) and triggering of cytotoxicity, allowing NK cells to act in
innate responses to tumors, transplants and infectious agents. A main
challenge is to understand critical receptor-ligand interactions during
NK cell development and education as well as in different pathophysiological situations. This is explored in collaborations with other MTC
scientists Petter Höglund, Maria Johansson, Björn Önfelt, Louise
Berg and Jonas Sundbäck (see their respective project reports).
In addition, we have collaborated with E Carbone and F Colucci to
identify NK receptors involved in recognition of primary and metastatic melanoma. As to autoimmunity, we have shown that the NKG2A
receptor is a key regulator of cytokine production by NK cells in the
synovial fluid of rheumatoid arthritis patients. Through M Uhlin,
we have collaborated with the “Centre for Allogeneic Bone Marrow
Transplantation” (KI Huddinge), in attempts to optimize expansion
of T cells of therapeutic interest. A transdisciplinary collaboration led
by S Cullheim at Department of Neuroscience, KI, has revealed a novel
role for MHC class I molecules in synaptic plasticity and regeneration
of motor neurons after axon injury, exerting influence on the afferent
input as well as on restoration of function at the motor end plate.
Selected publications
Selected publications
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Forsell MNE, Dey B, Mörner A, Svehla K, O’dell S, Högerkorp
C-M, Voss G, Thorstensson R, Shaw G, Mascola J, Karlsson Hedestam GB and Wyatt RT. B cell recognition of the conserved HIV-1
co-receptor binding site is altered by endogenous primate CD4.
PLoS Pathog 4:e1000171, 2008.
Mörner A, I Douagi A, Forsell M N E, Sundling C, Dosenovic P,
O’dell S, Dey B, Kwong P, Voss G, Thorstensson R, Mascola J, Wyatt
R and Karlsson Hedestam GB. HIV-1 Env-trimer immunization
of macaques and impact of priming with viral vector or stabilized
core protein. Virol 83, 540, 2009.
Dosenovic P, Chakrabarti B, Soldemo M, Douagi I, Forsell MNE,
Li Y, Phogat A, Paulie S, Hoxie J, Wyatt R T and Karlsson Hedestam G B. Selective expansion of HIV-1 Env-specific B cell subsets
recognizing distinct structural elements following immunization.
Immunol 183, 3373, 2009.
Lesion induced increase of motoneuron
MHC class Ia, with
predominant expression
in regenerating axons
and at re- innervated
neuromuscular junctions
(NMJ). Figure shows
colocalization between
H2-Db (middle panel)
and presynaptic markers
(left panel) in normal and
reinnervated NMJs (from
Thams et al., 2009).
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de Matos CT, Berg L, Michaëlsson J, Felländer-Tsai L, Kärre K and
Söderström K. Activating and inhibitory receptors on synovial
fluid natural killer cells of arthritis patients: role of CD94/NKG2A
in control of cytokine secretion. Immunology 122, 291, 2007.
Lakshmikanth T, Burke S, Ali TH, Kimpfler S, Ursini F, Ruggeri
L, Capanni M, Umansky V, Paschen A, Sucker A, Pende D, Groh
V, Biassoni R, Höglund P, Kato M, Shibuya K, Schadendorf D,
Anichini A, Ferrone S, Velardi A, Kärre K, Shibuya A, Carbone E
and Colucci F. NCRs and DNAM-1 mediate NK cell recognition
and lysis of human and mouse melanoma cell lines in vitro and in
vivo. J Clin Invest 119, 1251, 2009.
Thams S, Brodin P, Plantman S, Saxelin R, Kärre K and Cullheim
S. Classical major histocompatibility complex class I molecules in
motoneurons: new actors at the neuromuscular junction. J Neurosci 29,13503, 2009.
Department of Microbiology, Tumor and Cell Biology – MTC
33
Research Reports Immunobiology
Vaccines and
Immunity
We are developing new vaccines against infectious disease and cancer. In this context, we
study innate and acquired immune responses
to various types of vaccines. We also build
Peter Liljeström
vehicles for use in vaccination and one of our
[email protected]
most central vaccine technology is in-house
developed alphavirus replicon system based
on Semliki Forest virus (SFV), our working horse for many years.
Using SFV as model we have studied how viruses interact with the host
cell and how they trigger innate immune responses. Using this knowledge we are in a position to know how vaccines based on the SFV platform work and what kind of immune responses are obtained.
We have also studied how additional innate receptor ligands can be
incorporated in the SFV platform
for driving the immune response
in certain directions as for example to enhance the humoral (antibody) response. We have compared a
number of vaccine technologies in an Display of the 4E10 peptide
effort to expand our understanding on epitope of the HIV-1 gp41 envehow different vaccines induce protec- lope protein (picture: Daniel
Johansson).
tive immune responses. We have also
combined different vaccine technologies in so called prime-boost regimens and found that such approaches modulate the responses in various
ways. Our work has also included in-depth analysis of T cell responses in
terms of quality and longevity with the goal of defining immune correlates for protection. We have found that there are significant differences
between viral vector vaccines.
In the area of HIV/AIDS, we have participated in the development of novel vaccine candidates, some of which have already entered
several clinical trials. Indeed, a DNA prime – NYVAC (poxvirus) boost
vaccination regimen proved to be very promising and is now scheduled to move into larger clinical testing. In our future work, we will
study the immune responses to a variety of vaccine candidates and
characterize in detail the responses in terms of quality, memory and
recall capacity. In particular, we want to define how the initial innate
responses in each case orchestrate the acquired immune responses.
Visualizing
protective immune
responses
Cellular CD4+ and CD8+ T cell responses
are protective against tuberculosis and
contribute to long-term containment of
Mycobacterium tuberculosis (Mtb) infection. Definition of Mtb specific CD4+ and
Mark Maeurer
CD8+ T cell response would be desirable
[email protected]
to gauge protective immune responses in
individuals i) vaccinated with BCG and/or
ii) exposed to Mtb, but who are protected (latent TB). Screening of the
4500 proteins from Mtb with T cells from clinically well defined individuals (active BCG vaccination, latent TB = protected, active TB) is
not feasible, yet screening for serum reactivity (IgG and IgA) is possible. We predicted that serum IgG and IgA responses are indicative of
T cell responses. B-cell and CD4+ T cell responses are closely related:
Screening of high-content Mtb chips with serum from clinically
very well defined populations enabled us to select promising target
antigens to gauge for clinically and biologically relevant biomarkers associated with Mtb infection and to identify rational vaccine
targets. Humoral Mtb targets were screened for CD8+ and CD4+ T
cell targets using peptide arrays and soluble recombinant MHC class
II molecules. A particular feature of long-lasting memory CD8+ T cell
responses turned out to be the expression of the homodimer CD8aa
– and the dependency on IL-7-signaling. IL-7-signaling can be significantly impaired in long-lasting chronic infections and be altered by
tissue-specific expression of IL-7 isoforms (alternative splicing). Of
great interest, IL-7 and an alternatively spliced isoform showed effects
on differentiation of human neuronal stem cells. We are currently
testing promising Mtb vaccine candidates and evaluate IL-7 isoforms
to augment long-lived cellular protective immune responses.
Selected publications
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Näslund TI, Uyttenhove C, Nordström EK, Colau D, Warnier
G, Jondal M, Van den Eynde BJ and Liljeström P. Comparative
prime-boost vaccinations using Semliki Forest virus, adenovirus
and ALVAC vectors demonstrate differences in the generation of a
protective central memory CTL response against the P815 tumor.
J Immunol 178, 6761, 2007.
Harari A, Bart PA, Stöhr W, Tapia G, Garcia M, Medjitna-Rais E,
Burnet S, Cellerai C, Erlwein O, Barber T, Moog C, Liljeström P,
Wagner R, Wolf H, Kraehenbuhl JP, Esteban M, Heeney J, Frachette
MJ, Tartaglia J, McCormack S, Babiker A, Weber J and Pantaleo
G. An HIV-1 clade C DNA prime, NYVAC boost vaccine regimen
induces reliable, polyfunctional and long-lasting T cell responses. J
Exp Med 205, 63, 2008.
Mooij P, Balla-Jhagjhoorsingh SS, Beenhakker N, van Haaften P,
Baak I, Nieuwenhuis IG, Heidari S, Wolf H, Frachette MJ, Bieler K,
Sheppard N, Harari A, Bart PA, Liljeström P, Wagner R, Pantaleo G
and Heeney JL. Comparison of human and rhesus macaque T cell
responses elicited by boosting with NYVAC encoding human immunodeficiency virus type 1 clade C immunogens. J Virol 83, 5881, 2009.
Department of Microbiology, Tumor and Cell Biology – MTC
Humoral recognition pattern analysis using a peptide microarray chip. Pattern obtained in serum from individuals who received a 5 component pertussis vaccine (left, top panel) vs placebo (left, bottom) defined by epitope
recognition intensities. Differences are visualized to the right: recognition
hotspots induced by vaccination.
Selected publications
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Weichold FF, Mueller S, Kortsik C, Hitzler WE, Wulf M, Hone DM,
Sadoff JC and Maeurer M. Impact of MHC class I alleles on the M.
tuberculosis antigen-specific CD8+ T cell response in patients with
pulmonary tuberculosis. Genes Immun 8, 334, 2007.
Gaseitsiwe S, Valentine D, Ahmed R Mahdavifar S, Magalhaes I
Zerweck J, Schutkowski M, Gautherot E, Montero F, Ehrnst A,
Reilly M and Maeurer M. MHC class II molecule- peptide complex
analysis on a microarray chip. Clin Vaccine Immunol. 16, 567, 2009.
Moors M, Vudattu N K, Abel J, Kramer U, Rane L, Ulfig N, Seccatelli S, Fritsche E and Maeurer M. Interleukin-7 and Interleukin-7
splice variants differentiate human neural progenitor cells. Genes
Immun 11, 11, 2010.
Research Reports Immunobiology
Regulation of gene
expression during
viral infection
All viruses, regardless of their host
tropism, genome structure or any other
properties, must use components of the
host cell gene expression machinery in
order to propagate. This reliance has led
Gerald McInerney
to the development of an evolutionary
[email protected]
arms race between viruses and hosts.
Viral strategies have evolved to ensure
access to this machinery while host cells have developed means to
recognize viruses as foreign entities and to restrict their expression. In
turn, viruses have developed methods to counteract these restrictions
and to reduce the cells’ ability to communicate with its neighboring
cells.
Our work concerns the mechanisms of regulation of gene expression during the early stages of viral infections. Using Semliki Forest
virus (SFV) as a model, we are studying viral interactions with diverse
cellular pathways and systems in order to understand the biology of
the virus-infected cell.
SFV induces the transient formation of stress granules in the cytoplasm of infected cells (McInerney et al., 2005). These granules are
sites of mRNA triage which may play a role in the shut off of host cell
mRNA translation. We are currently investigating the mechanisms by
which these granules are assembled and disassembled and the nature
of their contribution to SFV cytotoxicity.
SFV infection induces the activation and nuclear localization of
type I interferon transcription factors IRF-3 and NFκB (see figure).
One of the SFV non-structural proteins, nsp2, also localizes to the
nucleus of infected cells and acts to inhibit the expression of type I
interferon (Breakwell, et al., 2007). Our recent work is aimed at
understanding how this viral protein inhibits host cell innate immune
signaling.
In addition to these projects, we work closely with other members
of the Gunilla Karlsson Hedestam group towards our common goal
of a deeper understanding the cell biology of virus infection and the
generation of virus-specific immune responses.
Semliki Forest virus
dependent activation
of interferon regulatory factor 3: IRF3
staining (red) shows
that the protein is activated by translocation
to the nucleus only in
the SFV-EGFP infected cells (green).
Altered T cell
homeostasis in
HIV-1 infection
T lymphocytes show high sensitivity to spontaneous and death receptor-induced apoptosis following HIV-1 infection. The death
receptor molecule Fas may play an important
role in T cell depletion as Fas expression on
T cells, the levels of FasL and sensitivity to
activation-induced apoptosis are all increased
in parallel with disease progression in HIV-1
Bence Rethi
[email protected]
infected patients.
In our studies we analyzed how the anti-apoptotic and T cell stimulatory cytokine IL-7 modulates the function of Fas on T cells. IL-7 is
particularly interesting in HIV infection as the concentration of this
cytokine is increasing in parallel with CD4+ T cell depletion, possibly
as a feedback mechanism that counteracts further T cell loss.
We demonstrated that IL-7 upregulates Fas expression on naïve
and memory T cells and induces a cytoskeleton-dependent Fas polarization on the cell surface. T cells, when treated with IL-7, undergo
apoptosis upon Fas cross-linking. IL-7 concentrations in the serum
of HIV-1 infected patients correlated with levels of Fas expression
on T cells and the sensitivity to Fas-mediated apoptosis. Fas, on the
other hand, can stimulate
proliferation when triggered on the surface of
sub-optimally activated
T cells. We demonstrated
that T cells of HIV-infected individuals are characterized by high sensitivity
to Fas-mediated costimulatory signals. Our results
suggest a scenario where
high IL-7 doses increase
Fas expression and induce
Fas membrane polariza- Regulation of Fas-mediated T cell apoptosis by IL-7 in lymphopenic conditions.
tion, an expression pattern
reminiscent of chronically
activated T cells. Triggering of Fas molecules may lead to increased
apoptosis of non-activated T cells whereas Fas signals, when induced
in weakly activated T cells, may rather contribute to proliferation.
T cell modulation by IL-7 gained further importance in light of
recent advances in IL-7 therapy applied to lymphopenic patients. We
continue to study how IL-7 modulates T cells functions with special
emphasis on T cell-B cell communication.
Selected publications
Selected publications
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Douagi I, McInerney GM, Hidmark AS, Miriallis V, Johansen K,
Svensson L and Karlsson Hedestam GB. Role of interferon regulatory factor 3 in type I interferon responses in rotavirus-infected
dendritic cells and fibroblasts. J Virol 81, 2758, 2007.
Breakwell L, Dosenovic P, Karlsson Hedestam GB, D’Amato M,
Liljeström P, Fazakerley J and McInerney GM. Semliki Forest virus
nonstructural protein 2 is involved in suppression of the type I
interferon response. J Virol 81, 8677, 2007.
McInerney G and Karlsson Hedestam G. Direct cleavage, proteasomal degradation and sequestration: Three mechanisms of viral
subversion of the type I interferon response. J Innate Immun 1,
599, 2009.
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Fluur C, De Milito A, Fry TJ, Vivar N, Eidsmo L, Atlas A, Federici C, Matarrese P, Logozzi M, Rajnavölgyi E, Mackall CL, Fais S,
Chiodi F and Rethi B. Potential role for IL-7 in Fas-mediated T cell
apoptosis during HIV infection. J Immunol 178, 5340, 2007.
Fluur C, Rethi B, Thang PH, Vivar N, Mowafi F, Lopalco L, Foppa
CU, Karlsson A, Tambussi G and Chiodi F. Relationship between
serum IL-7 concentrations and lymphopenia upon different levels
of HIV immune control. AIDS 21, 1048, 2007.
Rethi B, Vivar N, Sammicheli S and Chiodi F. Limited efficiency
of endogenous interleukin-7 levels in T cell reconstitution during
HIV-1 infection: will exogenous interleukin-7 therapy work?
AIDS 23, 745, 2009.
Department of Microbiology, Tumor and Cell Biology – MTC
35
Research Reports Immunobiology
Studies on the control
of bacterial and
parasitic infections
The global threats to public health
posed by the human pathogen
Mycobacterium tuberculosis (Mtb)
the causative agent of pulmonary
tuberculosis (TB) require our immeMarkus Sköld
diate attention. Our goal is to better
[email protected]
understand protective immunity
during pulmonary TB focusing on
the in vivo role of monocyte-derived cells. Monocytes are formed in
the bone marrow and are constantly released into circulation during
steady-state and inflammatory conditions. Using a newly developed monocyte adoptive transfer model, we have shown that many
of the macrophage and dendritic cell (DC) subsets that appear in
Mtb infected lung tissue and pulmonary lymph nodes (PLN) are
monocyte-derived. Macrophages and DC are well suited to influence multiple aspects of host immunity during pulmonary TB. Our
data show that activated monocyte-derived macrophages recruited to
Mtb infected lungs induce expression of enzymes important for their
bactericidal activity. In contrast, monocytes recruited to infected PLN
draining the lung tissue acquire a mature DC phenotype to help initiate the adaptive immune response by priming naïve T lymphocytes.
To study the host immune response during pulmonary TB
we have, together with our colleagues at the Swedish Institute for
Infectious Disease
Control, established
a murine low dose
(20-200 colony forming units) nose-only
Mtb aerosol infection
model in a Biosafety
Level-3 animal facility. This system
mimics the natural
Rod-shaped acid-fast bacilli (stained red)
route of infection and
detectable in lung tissue sections from infected
mice infected with M. tuberculosis.
allows us to determine how monocyte differentiation and function correlates with Mtb susceptibility.
Better understanding of susceptibility to Mtb in the mouse model
may provide valuable insights into susceptibility in humans. Because
Mtb is the largest killer of people co-infected with human immunodeficiency virus-1 (HIV-1), we are currently extending our studies on
monocyte function and the pathogen-specific T cell response in vivo
to a unique murine model of Mtb/HIV co-infection.
Selected publications
Selected publications
m­echanisms.
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Monocyte differentiation
and function during
pulmonary tuberculosis
We study the role and regulation of
IFN-γ secretion and responses during
intracellular chlamydial and mycobacterial infections, focusing on the role
of “suppressor of cytokine signaling 1”,
Martin Rottenberg
a protein that inhibits IFN-γ signaling
Hans Wigzell
(Figure). We found that bacterial innate
[email protected]
receptor agonists induce SOCS1 which
hampers bacterial growth control in
macrophages and bacterial antigen presentation. SOCS1 inhibited not
only IFN-γ-mediated responses but also the secretion of IFN-γ itself.
In vivo, SOCS1 hampered Mycobacterium tuberculosis and Chlamydia
growth control, but protected the host from severe pulmonary inflammation. We propose that targeting SOCS1 in defined cellular populations will contribute to protection against these infections.
Immunodeficient mice inoculated with CD34+ human hematopoietic stem cells generate human myeloid and lymphoid cell
populations de novo. We have used such a humanized mouse model
to study human immune responses to mycobacteria. We found that
human T cells differentiate into effector memory cells, migrate into
the lung and secrete human IFN-γ in humanized mice infected with
mycobacteria BCG. Human T cells were able to protect mice infected
with BCG. The generation of human adaptive immune responses and
the role of different T cell populations in the control of mycobacterial
infections in humanized mice is currently being studied.
There is an urgent need to substitute the highly toxic compounds
still in use for treatment of the encephalitic stage of African trypanosomiasis, a disease caused by infection with the protozoan Trypanosoma brucei. Trypanosomes do not synthesize their own purines.
Cordycepin was selected from a parasite viability screening of a library
of nucleoside analogues. When administered together with the adenosine deaminase inhibitor deoxycoformycin, cordycepin cured mice
even after parasites had penetrated into the brain. Cordycepin is cheap,
non-toxic, and has been tested in clinical trials in leukemic patients. Our
studies support
the clinical testing of treatment
with cordycepin
and deoxycoformycin
against
SOCS1 is induced by bacterial innate recephuman African
tor ligands and hampers secretion of IFN-γ, and
trypanosomiasis.
thereby the induction of IFN-γ-dependent bacterial
Yang T, Stark P, Janik K, Wigzell H and Rottenberg ME. SOCS-1
protects against Chlamydia pneumoniae-induced lethal inflammation but hampers effective bacterial clearance. J Immunol 180,
4040, 2008.
Vodnala SK, Ferella M, Lundén-Miguel H, Betha E, van Reet N,
Amin DN, Oberg B, Andersson B, Kristensson K, Wigzell H and
Rottenberg ME. Preclinical assessment of the treatment of secondstage african trypanosomiasis with cordycepin and deoxycoformycin. PLoS Negl Trop Dis 3:e 495, 2009.
Claes F, Vodnala SK, van Reet N, Boucher N, Lunden-Miguel H,
Baltz T, Goddeeris BM, Buscher P and Rottenberg ME. Bioluminescent imaging of Trypanosoma brucei shows preferential testis
dissemination which may hamper drug efficacy in sleeping sickness. PLoS Negl Trop Dis 3,e 486, 2009.
Department of Microbiology, Tumor and Cell Biology – MTC
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Sköld M and Behar SM. Tuberculosis triggers a tissue dependent
program of differentiation and acquisition of effector functions by
circulating monocytes. J Immunol 181, 6349, 2008.
Kiener HP, Watts GFM, Cui Y, Wright J, Thornhill TS, Sköld M,
Behar SM, Niederreiter B, Lu J, Cernadas MM, Coyle AJ, Sims G,
Smolen J, Warman ML, Brenner MB and Lee DM. Synovial fibroblasts self-direct multicellular lining architecture and synthetic
function in three-dimensional organ culture. Arthritis Rheum 62,
742, 2010.
Sada-Ovalle I, Sköld M, Tian T, Besra GS and Behar SM, a-Galactosylceramide as a therapeutic agent for pulmonary Mycobacterium tuberculosis infection. Am J Respir Crit Care Med, In press,
2010.
Research Reports Immunobiology
Regulation of anti-bacterial
responses of macrophages
by the receptor TRAPC
We have cloned and are studying
the function of a novel triggering receptor expressed on macrophages, dendritic cells and B cells,
i.e. antigen presenting cells, which
Jonas Sundbäck
we have named TRAPC. TRAPC
[email protected]
belongs to the TREM family of
molecules, which contains family
members involved in activating macrophages during bacterial infections, are expressed on osteoclasts and involved in bone resorption,
and are involved in modulating the function of microglial cells. The
TRAPC receptor associates with the adaptor signaling molecule
DAP12, and its expression is upregulated by bacterially derived LPS.
Cross-linking of TRAPC on macrophages and dendritic cell activates
the cells to produce nitric oxide, and protects macrophages from
bacterial infection, suggesting a role for the receptor in early defense
from bacteria.
It has been shown that TLR signaling can be inhibited by DAP12
signaling. The TRAPC receptor can balance signals from TLR receptors in order to fine-tune the receptor signaling in presence or absence
of a bacterial infection. One way to increase the understanding of the
function of the receptor is to identifying the ligand for TRAPC. We
have constructed a TRAPC-Fc fusion protein to use in the search of
the ligand. This fusion protein bound a potential ligand on macrophage cell lines, and will continue by immunoprecipitating and
protein sequencing to identify the ligand. Given the ability of TRAPC
to balance LPS mediated activation of macrophages in an experimental setting of bacterial infection, it would be interesting to further
study the role of the receptor using in vivo models, and to extend the
studies to other cells where it is expressed.
Genomic localization of TRAPC on mouse chromosome 17.
Visualizing
immune cell
interactions
The primary focus of our research is to understand the processes in which natural killer
(NK) cells and T cells communicate with
and eliminate cells that are potentially harmful to the body. The heterogeneity regarding
Björn Önfelt
the receptor expression patterns and effector
[email protected]
functions of the target cell populations makes
these phenomena utterly complex. Most
immunological setups in this field focus on readouts based on the
average of the cell population. Thus far we have pursued the goal of
developing new assays that can be used in academic research as well as
the pharmaceutical and health care industry. We focus on approaches
that provide the means to study the behavior of single cells and small
subpopulations of cells. In the future this will enable us to effectively
study how individual NK cells respond to different conditions.
Cell mediated immunity by NK cells is achieved by the formation
of an immune synapse. This is a highly organized subcellular interface
between the NK and the target cell involving a number of communication processes leading to secretion of
cytotoxic molecules
and killing of the target cell. To address
heterogeneity of NK
cell populations we
have developed two
m i c ro ch i p - b a s e d
approaches for high
Schematic of a microchip made in silicon-glass
resolution imaging
mounted in a custom made holder. Black line
of individual NK
shows a trajectory of a NK cell (blue) interacting
cells or NK cell-tarwith tumor target cells (yellow).
get-cell conjugates
over extended periods of time. The cells are restricted in space by miniature wells organized in an array format or an ultrasonic cage, which
promotes and facilitates studies of cell-cell interactions.
In order to characterize the intermittent movements of
lymphocytes we implement a method that has previously been used
in single molecule tracking studies. That makes it possible to automatically detect and quantify the periods of transient migration arrest
that is typical for these cells. The method allows quick assessment and
quantification of altered migration properties of cells during e.g. drug
treatments and infections.
Selected publications
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Selected publications
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Sandberg J K, Franksson L, Sundbäck J, Michaëlsson J, Petersson
M, Achour A, Wallin R P, Sherman N E, Bergman T, Jörnvall H,
Hunt D F, Kiessling R and Kärre K. T cell tolerance based on avidity
thresholds rather than complete deletion allows maintenance of
maximal repertoire diversity. J Immunol 165, 25, 2000.
Sundbäck J, Achour A, Michaëlsson J, Lindström H and Kärre K.
NK cell inhibitory receptor Ly-49C residues involved in MHC class
I binding. J Immunol 168, 793, 2002.
Chen, L, Sundbäck J, Olofsson S and Jondal M. Interference with
O-glycosylation in RMA lymphoma cells leads to a reduced in vivo
growth of the tumor. Int J Cancer 119, 1495, 2006.
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Sowinski S, Jolly C, Berninghausen O, Purbhoo M A, Chauveau A,
Kohler K, Oddos S, Eissmann P, Brodsky F M, Hopkins C, Önfelt
B, Sattentau Q and Davis DM. Membrane nanotubes physically
connect T cells over long distances presenting a novel route for
HIV-1 transmission. Nat Cell Biol 10, 211, 2008.
Benninger R K P, Vanherberghen B, Young S, Taner S B, Culley F
J, Schnyder T, Neil M A, Wüstner D, French P M, Davis DM and
Önfelt B. Live cell linear dichroism imaging reveals extensive
membrane ruffling within the docking structure of natural killer
cell immune synapses. Biophy J 96, L13, 2009.
Manneberg O, Vanherberghen B, Önfelt B and Wiklund M. Flowfree transport of cells in microchannels by frequency-modulated
ultrasound. Lab on a Chip 9, 833, 2009.
Department of Microbiology, Tumor and Cell Biology – MTC
37
Research Reports Infection Biology
Infection Biology Research
– broad, yet clearly focused to meet the emerging challenges ahead
Infection biology includes all the disciplines of classical medical microbiology. Yet,
medical microbiology of today tends to study
microbes in their close relation to the host,
host responses and in the perspective of the
co-evolution of microbes with their hosts.
Thus, apart from describing the pathogens
themselves, infection biology also deals with
events such as microbe-induced host cell
apoptosis, signal transduction, actin polymerization and cell cycle control.
As a result, infection biology has experienced a strong development into a multi-
38
and often a collaborative tradition at MTC.
The fact that many research groups working within the subject area are involved in
a number of national and international
collaborations and the functional closeness
to the Swedish Institute for Infectious Disease
Control, all adds to the diversity and resources.
disciplinary research field with obvious
connections with immunology during the
last decade, including innate immunity and
inflammation, vaccinology and even carcinogenesis. Not surprisingly therefore, the
research area of infection biology has found
a rather fruitful platform for establishing
itself within MTC. Topics covering bacterial,
parasitic and viral pathogenesis, the activity
of virulence factors, the formation of bacterial biofilms, molecular basis for resistance
to antimicrobial agents, host responses and
vaccine development have a long-standing
Infection biology research at MTC
Despite modern hygiene standards and treatment regimens, infectious diseases still pose
a major threat to human health. Classical
menaces such as malaria, pneumonia and
Hannah Akuffo
Jan Albert
Tobias Allander
Annelie Brauner
Lars Engstrand
Anneka Ehrnst
Jan-Ingmar Flock
Marta Granström
Birgitta Henriques
Normark
Göran Kronvall
Ewert Linder
Åke Lundkvist
Lars Magnius
Tore Midvedt
Roland Möllby
Elisabeth Norin
Staffan Normark
Mikael Rhen
Ute Römling
Mats Wahlgren
Britta Wahren
Project leaders:
Mats Andersson
Andreas Bråve
Fredric Carlsson
Qijun Chen
Christian Giske
Maria Isaguliants
Department of Microbiology, Tumor and Cell Biology – MTC
Marianne Jansson
Annika Karlsson
Jonas Klingström
Öjar Melefors
Sarah Palmer
Kristina Persson
Laura Plant
Katrin Pütsep
Research Reports Infection Biology
tuberculosis, as well as diarrhoeal diseases still
prevail, while mankind is exposed to additional infectious entanglements such as HIV,
emerging variants of influenza viruses and
the rapid development of resistance to antimicrobials.
As a whole, the coverage of the infection
biology research at MTC is fairly broad, yet
clearly focused to meet the emerging challenges ahead.
A major focus of the bacteriologists at
MTC is to understand how bacteria interact
with the epithelial linings and professional
phagocytic cells, and how these interactions
affect the concomitant host response and the
infection pathogenesis. Apart from defining
the molecular basis of bacterial virulence,
such studies have revealed microbial components suitable for vaccine development and
pharmaceutical inhibitors that can block
virulence factor activity.
Another central line of research concerns
the formation of bacterial biofilms, and role
of novel second messengers in the regulation
A particular interest is to characterize
of biofilm formation. Furthermore, research
many of the variable surface adhesins of the
is conducted on molecular epidemiology
parasite in terms of their antigenicity and
and to understand the origin and spread of
genetic regulation.
antimicrobial resistance
Studies on the human
amongst bacteria. MTC
immunodeficiency
virus
“Infection biology has
also conducts research on
(HIV)
and
the
Epstein-Barr
during the last decade
a theme called biomedivirus (EBV) continue to be
experienced a strong
cal ecology that in part
strong at MTC. Regarding
development into a multiinvestigates the role of the
HIV, the research covers
disciplinary research field
normal flora in human
clinical
epidemiology,
with obvious c
­ onnections
health and disease.
genome
variability,
viral
with immunology, including
evolution,
molecular
virolinnate immunity and inflamogy and vaccine developCreate vaccines
mation, ­vaccinology and
ment using a number of
MTC houses a strong
even carcinogenesis.”
approaches.
tradition in parasitology,
Other lines of interest include studies
and particularly in research devoted to Plason emerging viruses such as alphaviruses,
modium falciparum. Here, again, the ambidengue virus and new influenza virus varition is to define the parasite components
ants, as well as the use of Puumala virus as a
that enable the completion and success of
model to elucidate the infection pathogenesis
the replication cycle in man in order to create
of hemorrhagic fevers.
vaccines or chemical compounds that could
prevent or relieve states severe of malaria.
Mikael Rhen
Department of Microbiology, Tumor and Cell Biology – MTC
39
Research Reports Infection Biology
Molecular analyses of
host responses following
Leishmania infection
Leishmaniasis is a widely distributed disease affecting approximately 12 million people worldwide. The focus of our work is on
understanding natural killer (NK)
Hannah Akuffo
cell interactions in the early phase of
[email protected]
Leishmania infection, mechanisms
[email protected]
of ulcer formation during Cutaneous leishmanisis (CL), and the
effect of helminth infection on host response to vaccination against
tuberculosis with the Mycobacterium bovis Bacillus Calmette-Guérin
(BCG). We confirmed that promastigotes, the developmental form of
Leishmania parasites transmitted by the vector, induce cytokine secretion in NK cells (Lieke et al., 2008). Paradoxically, as a rapid consequence of this interaction, NK cell receptors show suppressive expression. We implicated the major surface molecules of promastigotes,
glycoprotein (gp) 63, in these suppressive effects. Currently, we are
following up the effects of Leishmania on NK cells by in vivo analysis
by taking advantage of a clinical trial in Iran, using Leishmania major
parasites for live vaccination.
Mechanisms of ulcer formation remain unclear. We reported that
Fas/FasL interactions are altered during CL and that keratinocyte
death through apoptosis may play a pathogenic role for skin ulceration (Eidsmo et al., 2007). We proposed, using keratinocytes incubated with supernatants from L. major-infected cells that activation
of Fas and TRAIL apoptosis pathways, in the presence of inflammatory mediators at the site of infection, leads to tissue destruction and
ulceration during CL. We furthermore studied the expression of FasL
and TRAIL in ulcerative and
non-ulcerative CL and found
higher FasL and TRAIL
expression in ulcerative
compared to non-ulcerative
CL. These findings correlated
with inhibition of ulcerative
processes in a murine CL
model following FasL and
TRAIL neutralization.
Currently, the only vaccine
available against tuberculosis
Leishmania promastigote (red arrow)
is BCG which has variable
meets natural killer cell (blue arrow).
Picture by Thorsten Lieke.
efficacy against pulmonary
tuberculosis. Work in our
group showed the chronic worm infection reduces the immunogenicity of BCG in humans which was associated with increased TGF-β
production, but not with enhanced Th2 immune response.
Selected publications
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Eidsmo L, Fluur C, Rethi B, Eriksson Ygberg S, Ruffin N, De
Milito A, Akuffo H and Chiodi F. FasL and TRAIL induce epidermal apoptosis and skin ulceration upon exposure to Leishmania
major. Am J Pathol 170, 227, 2007.
Lieke T, Nylén S, Eidsmo L, McMaster WR, Mohammadi AM,
Khamesipour A, Berg L and Akuffo H. Leishmania surface protein
gp63 binds directly to human natural killer cells and inhibits
proliferation. Clin Exp Immunol 153, 221, 2008.
Elias D, Britton S, Aseffa A, Engers H and Akuffo H. Poor immunogenicity of BCG in helminth infected population is associated with
increased in vitro TGF-b production. Vaccine 26, 3897, 2008.
Department of Microbiology, Tumor and Cell Biology – MTC
HIV genetic
variation
and evolution
The research in my group focuses on genetic
and biological variability of HIV as well as the
interactions between HIV and the immune
system. We study how HIV evades humoral and
cellular immunity. We also study the emergence
Jan Albert
and spread HIV variants that are resistant to
[email protected]
anti-retroviral drugs. We use molecular epidemiology to track the spread of HIV in Sweden
and abroad. We take part in several EU projects such as EuropeHIVResistance and CHAIN and other international projects
Our current research also focuses on the development and application of modern Bayesian methods for phylogenetic inference.
HIV prevention still relies solely on information campaigns and
other behavioral interventions. To guide prevention it is essential
to have correct and detailed information about the local and global
HIV epidemic. Phylodynamics is a rapidly developing interdisciplinary research field, which we use to document and time the HIV-1
outbreak among intravenous drug users in Stockholm, as well as the
entire HIV epidemics in Latvia and Honduras.
A second area of research is the application of massive parallel
pyrosequencing (454-sequencing) to dissect HIV evolution with a
previously unattainable resolution. 454-sequencing allows ultra-deep
sequencing and statistically meaningful quantification of low-abundance genetic variants,
such as rare sequence variants in an HIV quasispecies. Our preliminary data
indicate that previously
undetectable drug resistance can be detected down
to 0.06% mutation abundance. We have generated
more than one million
HIV sequences within this
project.
Finally, we take part
in an EU project called
NGIN which focuses on
neutralizing antibodies in
HIV infection and aims to
develop immunogens that
Example of modeling of HIV spread by
induce broad neutralization. sophisticated phylogenetic tree analyses.
Selected publications
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Skar H, Sylvan S, Hansson HB, Gustavsson O, Boman H, Albert
J and Leitner T. Multiple HIV-1 introductions into the Swedish
intravenous drug user population. Infect Genet Evol 8, 545, 2008.
Vercauteren J, Wensing AM, van de Vijver DA, Albert J, Balotta C,
Hamouda O, Kucherer C, Struck D, Schmit JC, Asjo B, Bruckova
M, Camacho RJ, Clotet B, Coughlan S, Grossman Z, Horban A,
Korn K, Kostrikis L, Nielsen C, Paraskevis D, Poljak M, Puchhammer-Stockl E, Riva C, Ruiz L, Salminen M, Schuurman R, Sonnerborg A, Stanekova D, Stanojevic M, Vandamme AM and Boucher
CA. Transmission of drug-resistant HIV-1 is stabilizing in Europe.
J Infect Dis 200, 1503, 2009.
Murillo W, de Rivera IL, Parham L, Jovel E, Palou E, Karlsson AC
and Albert J. Prevalence of drug resistance and importance of viral
load measurements in Honduran HIV-infected patients failing
antiretroviral treatment. HIV Med 11, 95, 2010.
Research Reports Infection Biology
Clinical virology
and virus
discovery
The group is based at the Department of
Clinical Microbiology at Karolinska University Hospital, and the research is focused on
developing efficient diagnostic methods for
virus infections. We have a special interest in
Tobias Allander
respiratory tract infections, but also study
[email protected]
gastroenteritis viruses and chronic virus
infections.
A main activity is identification of unknown viruses and clinical
characterization of newly discovered viruses. The group has identified two novel human viruses, human bocavirus (HBoV) and KI
polyomavirus (KIPyV). We have further established a relationship
between human bocavirus and acute respiratory disease in infants
and children. HBoV analysis is now routinely provided to health care.
Virus discovery is done by combining a specially designed virus
purification protocol with high-throughput sequencing and bioinformatics, in collaboration with the genome sequencing group at the
Department of Cell- and Molecular Biology, Karolinska Institutet. We
screen a wide variety of clinical samples for unknown viruses, from
upper respiratory tract secretions to tumors.
Virus infections impose a huge disease burden on humanity. Diagnosing infectious agents and searching for the cause for diseases with
unknown aetiology is fundamental for many aspects of clinical medicine.
A molecular diagnostic platform for daily rapid detection of 15 respiratory
viruses
Defending the uroepithelium
– new strategies fighting
urinary tract infections
The urinary tract functions in
close proximity to the outside
environment yet must remain
free of microbial colonization
to avoid disease. Recently, we
Annelie Brauner
demonstrated that epithelial cells
[email protected]
of the urinary tract up-regulate
the production of the human
antimicrobial peptide cathelicidin upon infection with uropathogenic Escherichia coli. Cathelicidin aggressively fends off bacteria,
thereby protecting the urinary tract from infection. However, if
bacteria succeed to adhere to the urinary bladder, they form biofilms,
which offer advantages to the bacteria, such as increased resistance
to antimicrobial substances, mechanical forces and to nutrient depletion. Biofilms are mainly built up by different extracellular matrix
components and substantially contribute to the complex interplay
between the host and the invading microorganism.
In spite of the effective defense system, urinary tract infection is
currently one of the 10 most common reasons for outpatient visits.
Up to 60 percent of all women develop the disease at least once in their
life, and of these, 25% have recurrent infections. Children suffer less
often from urinary tract infections but when the kidneys are involved
up to 40% get renal scarring.
Antibiotic-resistant bacteria are a growing and alarming problem
and therefore call for immediate action to develop new treatment
strategies. Our research interest focuses on patients with recurrent
urinary tract infections,
with special emphases on
the mechanism of action
between the microorganism, biofilm and innate
immunity. Particularly, we
concentrate on underlying mechanisms causing
recurrent infections and
possibilities to increase the
endogenous production of
cathelicidin and other novel
antimicrobial peptides. We
also focus on bacterial viruE. coli surrounded with curli fimbriae.
The bacteria were isolated directly from
lence factors enabling the
fresh urine of a patient with urinary tract
bacteria to withstand tradiinfection.
tional antibiotic treatment.
Selected publications
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Allander T, Andreasson K, Gupta S, Bjerkner A, Bogdanovic G,
Persson MA, Dalianis T, Ramqvist T and Andersson B. Identification of a third human polyomavirus. J Virol 81, 4130, 2007.
Tiveljung-Lindell A, Rotzén-Östlund M, Gupta S, Ullstrand R,
Grillner L, Zweygberg-Wirgart B and Allander T. Development
and implementation of a molecular diagnostic platform for daily
rapid detection of 15 respiratory viruses. J Med Virol 81, 167, 2009.
Goh S, Lindau C, Tiveljung-Lindell A and Allander T. Merkel cell
polyomavirus in respiratory tract secretions. Emerg Infect Dis 15,
489, 2009.
Selected publications
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Chromek M and Brauner A. Antimicrobial mechanisms of the
urinary tract. J Mol Med 86, 37, 2008.
Lüthje P and Brauner A. Putative link between the virulence-associated gene fluA and fluoroquinolone resistance in uropathogenic
E. coli. J Clin Microbiol 48, 675, 2010.
Kai-Larsen Y, Lüthje P, Chromek, M, Peters V, Wang X, Holm Å,
Kádas L, Hedlund K-O, Johanson J, Chapman MR, Jacobson SH,
Ute Römling U, Agerberth B, Brauner A . Uropathogenic Escherichia coli modulates immune responses and its curli fimbriae interact with the antimicrobial peptide LL-37. Accepted PLoS Pathogens 2010.
Department of Microbiology, Tumor and Cell Biology – MTC
41
Research Reports Infection Biology
Immunization strategies against
HIV-1: immunogen design and
optimization of vaccine delivery
More than 25 years have
passed since the identification and isolation of Human
Immunodeficiency
Virus
type 1 (HIV-1). Although
Andreas Bråve
there is still no effective
[email protected]
vaccine available against the
virus, recent results from
a phase 3 trial in Thailand have indicated that protective immunity
can be achieved by immunization. Due to the intrinsic nature of the
virus, classical vaccine approaches, such as inactivated virus vaccines
or protein-based vaccines, have failed and researchers are forced to
turn to more novel vaccine-strategies, such as gene-based vaccination (e.g. DNA-vaccination). However, DNA vaccines have so far
elicited weaker immune responses in humans than anticipated and,
in most current clinical trials, plasmid vaccines are used primarily to
“prime” the immune system for a subsequent boost with recombinant
protein or a live recombinant attenuated viral vector, such as Modified Vaccinia virus Ankara (MVA) or Adenovirus.
Our vaccine strategy against
HIV-1 is to target multiple viral
antigens deriving from various
subtypes of the virus. The results
from our clinical trials, in which HIV
DNA plasmids are combined with a
recombinant MVA, are encouraging
with close to 100% of the volunteers
Bifunctional vaccine-specific
responding against HIV-1 antigens T cell responses as analyzed
after immunization. We are now by FluoroSpot. T cells isolated
developing the next generation of from DNA-immunized mice
were stimulated with vaccine
genetic HIV-1 immunogens and also antigen and respond by producfocusing on improving the delivery ing either IFN-g (green), IL-2
of the plasmid DNA vaccine by the (red) or both (yellow).
use of electroporation. In vivo-electroporation is the application of short pulses of electric current immediately after, and at the site of, an injection with a genetic vaccine. We
and others have shown that it is possible to significantly augment the
subsequent vaccine-specific immune responses using this method.
Now, the overall aim is to develop an immunization strategy that will
allow plasmid DNA to be used as a stand-alone vaccine modality.
Host-pathogen
interactions in
mycobacterial infection
One third of the world’s population is infected with Mycobacterium
tuberculosis (Mtb), which causes
around 2 million deaths annually.
To facilitate fundamental studies
Fredric Carlsson
of Mtb infection, safer and [email protected]
mentally more amenable species are
often used as models. Among these,
the closely related Mycobacterium marinum (Mmar) is used increasingly to study mycobacterial pathogenesis. Mmar is a pathogen of fish
and amphibians causing disease with many features of tuberculosis,
and is also able to infect immunocompetent humans where it induces
formation of dermal granulomas pathologically similar to those
formed in tuberculosis. Thus, Mmar and Mtb likely share mechanisms of inducing and maintaining infection.
The overall goal of our work is to significantly increase our understanding of how pathogenic mycobacteria interact with the host
immune system to promote disease. One major interest in our lab
is to understand how the mycobacterial Esx-1 (type VII) secretion
system interacts with the host inflammasome, an important part of
our innate immune response. Preliminary data indicate that Esx1-dependent activation of the inflammasome is host-detrimental
during Mmar infection, suggesting that activation of this inflammatory pathway in mycobacterial infection is a manifestation of bacterial
virulence rather than a manifestation of host response. Another focus
in our lab is to elucidate the host-pathogen interactions whereby
mycobacteria interfere with the onset and efficacy of the adaptive immune response in a novel mouse model of Mmar infection.
It is well known that Mtb delays the onset of the adaptive immune
response both in humans and in mice, but the underlying molecular
events have remained unknown. This likely is central to the difficulties in generating an effective vaccine and in harnessing the human
immune system for therapy. Our studies aim to provide fundamental
knowledge to answer this key question.
Bone marrow-derived
macrophages from
B6 mice infected with
M. marinum. Blue:
Macrophage nuclei
stained by Hoechst
stain. Green: Active
caspase-1 stained
by FAM-FLICA as a
marker for inflammasome activation upon
infection.
Selected publications
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Bråve A, Boberg A, Gudmundsdotter L, Rollman E, Hallermalm K,
Ljungberg K, Blomberg P, Stout R, Paulie S, Sandström E, Biberfeld
G, Earl P, Moss B, Cox JH and Wahren B. A new multi-clade DNA
prime/recombinant MVA boost vaccine induces broad and high
levels of HIV-1-specific CD8+ T cell and humoral responses in
mice. Mol Ther 15, 1724, 2007.
Sandström E, Nilsson C, Hejdeman B, Bråve A, Bratt G, Robb M,
Cox, J, VanCott T, Marovich M, Stout R, Bakari M, Kisali Pallangyo,
Ljungberg K, Moss B, Earl P, Michael N, Birx D, Mhalu F, Wahren
B and Biberfeld G. Broad immunogenicity of a multigene, multiclade HIV-1 DNA vaccine boosted with heterologous HIV-1
recombinant modified vaccinia virus Ankara. J Infect Dis 198,
1482, 2008.
Bråve A, Hallengärd D, Malm M, Blazevic V, Rollman E, Stanescu I
and Krohn K. Combining DNA technologies and different modes
of immunization for induction of humoral and cellular antiHIV-1 immune responses. Vaccine 27, 184, 2009.
Department of Microbiology, Tumor and Cell Biology – MTC
Selected publications
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McLaughlin B, Chon JS, MacGurn JA, Carlsson F, Cheng TL, Cox
JS and Brown EJ. A Mycobacterium ESX-1-secreted virulence
factor with unique requirements for export. PLoS Pathog 3, e105,
2007.
Carlsson F and Brown EJ. The art of making an exit. Science 323,
1678, 2009.
Carlsson F, Joshi SA, Rangell L and Brown EJ. Polar localization of
virulence-related Esx-1 secretion in Mycobacteria. PLoS Pathog 5,
e1000285, 2009.
Research Reports Infection Biology
Interactions of malaria
virulence factors and
the human host
The malaria parasite Plasmodium falciparum exposes various molecules at
the surface of infected erythrocytes.
Some of the surface-exposed molecules
are parasite virulence factors associQijun Chen
ated with severe malaria and immune
[email protected]
evasion. Our interest is to understand
the genetic regulation and biological
function of the virulence factors named P. falciparum erythrocyte
membrane protein 1(PfEMP1) and Antigen 332 (also called Pf332).
We have investigated the transcription and regulation of the
var2csa gene that encodes a PfEMP1 associated with placental
malaria. We established an approach combining real-time PCR allelic
discrimination and discriminative RNA-FISH to distinguish between
highly similar gene copies in P. falciparum parasites. We found that
duplicated var2csa variants are simultaneously transcribed both on
a population level, and, intriguingly, also in individual cells, with
nuclear co-localization of the active genes and corresponding transcripts. This indicates transcriptional functionality of the duplicated
genes, challenges the dogma of mutually exclusive var gene transcription and suggests mechanisms behind antigenic variation. Allelic
discrimination, gene or transcript localization by FISH not only
provide insights into transcriptional regulation of genes, but also
suggest that this approach could be used for further investigation of
genome dynamics and gene regulation.
Another recent focus has been on the molecule that mediates
erythrocyte invasion by the P. falciparum parasite. We found that
the Pf332 molecule harbors a Duffy-binding like (DBL). The DBL
domain of Pf332 is conserved among parasites isolated from different
regions in the world. Importantly, affinity purified human antibodies as well as mAb to the DBL domain significantly inhibit invasion,
indicating that the DBL domain plays a critical role for parasite invasion. The identification of the molecule paves a new way towards the
development of a malaria vaccine.
Characteristics of trans­
missible HIV-1 and
co-receptor use
Detection of var gene transcripts in single parasites by RNA-FISH.
HIV mother-to-child transmission (MTCT) is preventable
in industrialized countries, but
remains an obstacle in most
developing countries. Our
Anneka Ehrnst
research aims at improving the
[email protected]
prevention of mother-to-child
transmission also in these countries. We study the role of HIV-1 subtypes and phenotypes for viral
entry into cells and their role in transmission and disease progression.
MTCT of HIV-1 could be decreased in Northern Vietnam, after
introducing counseling, avoidance of breastfeeding, and anti-retroviral therapy (Ha et al., 2008), despite that women here usually escape
from follow-up programs due to fear. We also determined the phylogenetic origin of their HIV virus. Combining the sequences of biological phenotypes of HIV
isolates and those of molecular clones, we can pinpoint
the exact transmitted virus.
These are analyzed genetically and in neutralization
to determine crucial properties in transmission. We
also characterize co-receptor use of different subtypes
HIV-1 binds to CD4 and to one of the
and their corresponding
co-receptors, CCR5 or CXCR4, when
infecting a cell. The normal function of
similar or different amino
CD4 is to bind to MHC class II molecules
acids in the critical V3 loop
and the function of CCR5 and CXCR4 is
of the env gp120 gene and
to bind chemokines.
change of co-receptor use in
drug failure. An algorithm to determine co-receptor use is compared
to other biometric methods and biological determination.
A microchip analysis of HIV-1 peptides reacting with monomeric
molecules of the major histocompatibility complex (HLA) class
II (Gaseitsiwe et al., 2009) was developed, which confirmed several
already known peptides and identified several new ones.
Furthermore, we analyze the role of cytomegalovirus (CMV) in
terminal illness of AIDS patients. When CMV was localized in the
same area of the brain affected in Korsakoff ’s syndrome, which is ususlly caused by chronic alcoholism, Korsakoff ’s syndrome occurred in
the absence of alcohol abuse (Matell-Pirskanen et al., 2009). The role
of CMV in other central nervous system-related illnesses is under way,
such as a CMV-related Addison’s disease, a rare endocrine disorder
wherein the adrenal glands produce insufficient steroid hormones.
Selected publications
Selected publications
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Chen Q. The naturally acquired immunity in severe malaria and
its implication for a PfEMP1 based vaccine. Microb Infect 9, 777,
2007.
Moll K, Chêne A, Ribacke U, Kaneko O, Nilsson S, Winter G,
Haeggström M, Pan W, Berzins K, Wahlgren M and Chen Q. A
novel DBL-domain of the P. falciparum 332 molecule possibly
involved in erythrocyte adhesion. PLoS One 2, e477, 2007.
Brolin KJ, Ribacke U, Nilsson S, Ankarklev J, Moll K, Wahlgren M
and Chen Q. Simultaneous transcription of duplicated var2csa
gene copies in individual Plasmodium falciparum parasites.
Genome Biol 10, R117, 2009.
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Ha TTT, Tuan PL, Bao NH, Anh NM, Cam PD, Chiodi F and
Ehrnst A. Successful reduction of mother-to-child transmission of HIV-1 by nevirapin and non-breastfeeding in Hanoi and
Haiphon. Retrovirology 5, S1, P11, 2008.
Gaseitsiwe S, Valentini D, Ahmed R, Mahdavifar S, Magalhaes I,
Zerweck J, Schutkowski M, Gautherot E, Montero F, Ehrnst A,
Reilly M and Maeurer M. Major histocompatibility complex class
II molecule-human immunodeficiency virus peptide analysis
using a microarray chip. Clin Vaccine Immunol 16, 567, 2009.
Matell-Pirskanen R, Grützmeier S, Nennesmo I, Sandström E and
Ehrnst A. Impairment of short-term memory and Korskakoff
syndrome are common in AIDS patients with cytomegalovirus
encephalitis. Eur J Neurol 18, 48, 2009.
Department of Microbiology, Tumor and Cell Biology – MTC
43
Research Reports Infection Biology
The impact of the human
gastrointestinal microbiota
on health and disease
The work of my group aims at
creating a scientific environment
with an interdisciplinary approach
on the interface between epidemiology and the microbiological,
Lars Engstrand
immunological and genetic aspects
[email protected]
of chronic infectious diseases. One
focus is Helicobacter pylori infection and to answer the question: why do some people who are infected
become ill, while others do not? The long-term objective is to identify and characterize factors (both microbial and host related) that
determine or modify the risk of disease outcome observed in people
infected with this microorganism.
Humans represent complex microbial ecosystems, which are
postulated to contribute to both health maintenance and the development of chronic inflammatory diseases. Alterations in the human
gut microbiota have recently been associated with diseases such as
cancer, type II diabetes, inflammatory bowel disorder, allergy and
obesity. Determining the microbial composition in patients and
healthy controls may provide novel therapeutic targets. New highthroughput culture-independent molecular tools are now developed,
allowing the scientific community to characterize and understand the
microbial communities underpinning biological processes in unprecedented ways. Our ongoing projects have the potential to transform the situation of chronic inflammation in the gut from a purely
descriptive to a predictive framework, in which ecological principles
are integrated and exploited to biological systems and pathology. We
interact with a number of clinical colleagues, including epidemiologists to initiate extended population-based studies. We are currently
analyzing the human bacteriome in a number of cases and controls
from such studies. The results will then be transferred to studies of
host and microbial interactions, molecular epidemiology, biomarkers and prevention, clinical diagnostics and pathogenic mechanisms.
Heatmap of genera present in human stomachs. Red: Individuals with high grade corpus atrophy, white: Controls, blue:
Individuals treated with proton pump inhibitors and green:
Individuals with dominance of Helicobacter sequences.
­Visualized using heatmap with MultiExperimental Viewer 4_5.
Selected publications
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Andersson AF, Lindberg M, Jakobsson H, Bäckhed F, Nyrén P and
Engstrand L. Comparative analysis of human gut microbiota by
barcoded pyrosequencing. PLoS One 30, e2836, 2008.
Giannakis M, Chen SL, Karam SM, Engstrand L and Gordon JI.
Helicobacter pylori evolution during progression from chronic
atrophic gastritis to gastric cancer and its impact on gastric stem
cells. Proc Natl Acad Sci USA 105, 4358, 2008.
Dicksved J, Lindberg M, Rosenquist M, Enroth H, Jansson JK and
Engstrand L. Molecular characterization of the stomach microbiota in patients with gastric cancer and in controls. J Med Microbiol
58, 509, 2009.
Department of Microbiology, Tumor and Cell Biology – MTC
Vaccination against
streptococcal
infections
Recombinant surface proteins from streptococci and staphylococci can give rise to
protective antibodies. We have now taken
such studies beyond the stage of experimental mouse models and demonstrated
Jan-Ingmar Flock
vaccine efficacy on the target animal, the
[email protected]
horse.
Streptococcus equi is a serious pathogen causing strangles in the horse, an upper respiratory tract infection. The bacteria adhere to the tonsils and spread via the lymphatic
system. It is the most contagious and costly bacterial infection in the
horse and causes severe problems for the horse industry. No safe and
efficient vaccine is available.
In collaboration with a team at Swedish University of Agricultural
Sciences in Uppsala, we have vaccinated horses with various recombinant surface-located proteins from S. equi. Antigens to be tested
in the horse are first tested for protective efficacy in a mouse model
system. The vaccinated horses are subsequently experimentally
infected with S. equi and protection is monitored by determination
of body temperature, lymph node swelling, clinical symptoms, post
mortem and histopathological analysis. A protection level at around
80% has been achieved. The present work is focused on finding additional antigen combinations, identification of protective epitopes,
optimization of immunization route, dosage,
and duration of protection. Currently, antibodies from sera and
nasal washes from
vaccinated horses are
analyzed to further
understand the mechanism of protection. Like Elevated body temperature is a typical sign
many other bacteria, S. of strangles in the horse. The graph shows
how experimentally infected horses, which
equi has a very sophis- have been vaccinated with two different
ticated system for which combinations (A and B) of recombinant S.
immune evasion can equi proteins, are protected from infection.
Mean values from six horses in each group
here be studied in the are shown. The difference between vaccinatural host.
nated and non-vaccinated horses is statistiS. equi and Strep- cally significant (p<0.0001).
tococcus
pyogenes,
a human pathogen causing e.g. scarlet fever, are similar in genome
sequence, mode of infection and clinical symptoms. Some antigens
from S. equi used in the vaccine have orthologs in S. pyogenes, implying that the corresponding antigens from S. pyogenes could be used in
a vaccine against this pathogen.
Selected publications
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Waller A, Flock M, Smith K, Robinson C, Mitchell Z, Karlström
A, Lannergard J, Bergman R, Guss B and Flock J-I. Vaccination of
horses against strangles using recombinant antigens from Streptococcus equi. Vaccine 25, 3629, 2007.
Guss B, Flock M, Frykberg L, Waller A, Robinson C, Smith K
and Flock J-I. Getting to grips with strangles: an effective multicomponent recombinant vaccine for the protection of horses from
Streptococcus equi infection. PLoS Pathog 9, e1000584, 2009.
Hulting G, Flock M, Frykberg L, Lannergård, Flock J-I and Guss B.
Streptococcus equi encodes two specific IgG-degrading enzymes.
FEMS Microbiol Lett 298, 44, 2009.
Research Reports Infection Biology
Multidrug-resistant
Gram-negative bacilli
During the last three years my
research projects have concentrated
on three major topics, all related to
various aspects of multidrug-resistChristian Giske
ant Gram-negative bacilli:
[email protected]
The epidemiology of extendedspectrum
β-lactamase
(ESBL)
producing Enterobacteriaceae in Stockholm and factors influencing
fecal carriage of ESBL-producing Enterobacteriaceae. The first epidemiological project has revealed a mainly polyclonal epidemiology
with some minor strain clusters. A project where the dynamics of fecal
carriage of ESBL-producing Enterobacteriaceae are being evaluated
is ongoing, and currently around 80 patients have been included. A
total of 100 patients will finally be included, and will be followed for at
least one year. Strain and plasmid replicon typing will be conducted to
understand more about the dynamics of carriage, and data regarding
treatment and co-morbidities will be collected for all patients.
Multilocus sequence typing as a tool for identifying clones of
multidrug-resistant (MDR) Gram-negative bacilli with a high epidemic potential. The Klebsiella pneumoniae clone ST258, which is
responsible for the majority of cases of carbapenem-resistant Enterobacteriaceae spreading in
the US, and also in several European countries,
was first described by
our group. Also, we first
described clones of Pseudomonas aeruginosa that
have later been associated
with epidemic spread of
the metallo-β-lactamases
CC111 and CC235. Further, we were the first to
Multidrug-resistant Gram-negative bacilli –
detect the new metalloa current public health threat.
β-lactamase
NDM-1,
which is highly prevalent
in India.
Several projects related to laboratory methods of detection
of new β-lactamases are currently being carried out. Partly, these
projects explore phenotypic methods for confirming the presence of
β-lactamases, and partly criteria for screening for such β-lactamases.
Our results have attracted great interest both in Europe and in the US.
Selected publications
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Giske CG, Monnet DL, Cars O and Carmeli Y. Clinical and
economic impact of common multidrug-resistant gram-negative
bacilli. Antimicrob Agent Chemother 52, 813, 2008.
Kitchel B, Rasheed JK, Patel JB, Srinivasan A, Navon-Venezia S,
Carmeli Y, Brolund A and Giske CG. Molecular epidemiology
of KPC-producing Klebsiella pneumoniae isolates in the United
States: clonal expansion of multilocus sequence type 258. Antimicrob Agent Chemother 53, 3365, 2009.
Samuelsen Ø, Naseer U, Tofteland S, Skutlaberg DH, Onken
A, Hjetland R, Sundsfjord A and Giske CG. Emergence of clonally related Klebsiella pneumoniae isolates of sequence type 258
producing plasmid-mediated KPC carbapenemase in Norway and
Sweden. J Antimicrob Chemother 63, 654, 2009.
Helicobacter pylori
infection in Vietnam
Our group has studied Helicobacter
pylori infection in the context of a
developing country, i.e. Vietnam, where
peptic ulcer disease and gastric cancer
Marta Granström
are major public health issues. We could
[email protected]
show that for optimal performance of
the serological assay local strains had to
be used, a major problem for developing countries since the commercial kits available on the market have been made and evaluated for
developed countries. We could also show that H. pylori infection was
very common in Vietnam and significantly higher in Hanoi than in
a rural area, probably due to the higher level of crowding in the city.
The early infection in Vietnam also results on a high rate of peptic
ulcer disease in children and the need to find the optimal treatment
for them since there are no formal recommendations for eradication
treatment even in developed countries.
We conducted a large randomised, double-blind study to evaluate
two main treatment strategies, one with and one without clarithromycin. The outcome was rather low eradication rates of around 60%,
which is in contrast to the >80% obtained in developed countries.
Antibiotic resistance testing revealed that the cause for the low eradication rate was an unexpectedly high rate of clarithromycin resistance
of overall 50% in Vietnamese children, with the youngest children
having even higher rates. An important finding was that twice-daily
dosing of the proton-pump inhibitor significantly improved the
eradication rate in the clarithromycin-resistant strains.
The one year follow-up of children with previously eradicated
H. pylori showed a high reinfection rate, with the highest rate found
in the youngest age groups and then gradually decreasing until 9-11
years of age. The finding supports the notion of H. pylori infection
being acquired at an early age due to unknown mechanism(s) of
susceptibility.
Pharmacy outside a pediatric
hospital.
Selected publications
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Granström M, Lehours P, Bengtsson C and Megraud F. Diagnosis
of Helicobacter pylori. Helicobacter 13 Suppl 1, 7, 2008.
Nguyen TV, Bengtsson C, Nguyen GK and Granström M. Evaluation of a novel monoclonal-based antigen-in-stool enzyme
immunoassay (Premier Platinum HpSA PLUS) for diagnosis of
Helicobacter pylori infection in Vietnamese children. Helicobacter
13, 269, 2008.
Nguyen TV, Bengtsson C, Nguyen GK, Hoang TT, Phung DC,
Sörberg M and Granström M. Evaluation of two triple-therapy
regimens with metronidazole or clarithromycin for the eradication of H. pylori infection in Vietnamese children: a randomized,
double-blind clinical trial. Helicobacter 13, 550, 2008.
Department of Microbiology, Tumor and Cell Biology – MTC
45
Research Reports Infection Biology
Microbial vaccines as
targeted cargo, genetic
approach
Strong immune response against
primary resistance mutations can create
a bottle-neck to HIV-1 forbidding the
mutations to occur. Such response can
be induced by genes encoding drugMaria Isaguliants
resistant HIV enzymes. The idea is
[email protected]
unconventional and used only by our
group. These DNA-encoded immunogens are not particularly strong, but immunogenicity can be enhanced
if the enzymes are targeted to specific intracellular compartments. For
this, their genes are fused with sequences encoding intracellular sorting signals. Targeting to proteasome/MHC class I pathway has been
achieved by protein fusions to ornithine decarboxylase or its minimal
proteasome targeting moiety (ODCsig). Mice DNA-immunized with
chimeras of wild-type or drug-resistant reverse transcriptase (RT)
with ODCsig mounted an enhanced immune response as compared
to parental genes.
Targeting to lysosome/MHC class II pathway was achieved by
fusion to endosomal sorting signals. Potent cross-reactive immune
response against both the wild-type and drug-resistant HIV RT was
achieved after DNA immunization with RT chimeras fused to lysosome associated membrane protein I. Screening of immune response
in animals is done by in vivo
imaging (IVIS®) where the
decrease of expression of a
reporter gene co-injected with
DNA-immunogen due to killing of expressing cells by the
immune response is moniSubcellular localization of wild-type
(A) and drug-resistant (B) HIV-1
tored. We have, thus, pioneered
reverse transcriptase chimeras
the use of drug-resistant HIV
fused to lysosome associated memenzymes and their chimeras as
brane protein I (LAMP-1). Staining of
the cell nucleus in blue, LAMP-1 in
DNA-immunogens and demgreen, reverse transcriptase in red,
onstrated their potency. This
in field overlays.
motivates further progress
towards human immunotherapy that would prevent the evolvement
of drug resistant HIV.
Another vaccine project, carried on the hepatitis C virus model,
deals with increasing the efficacy of DNA immunization by applying different immunogen combinations, prime-boost regimens and
novel adjuvants.
Selected publications
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Petrakova N, Gudmundsdotter L, Yermalovich M, Belikov SV,
Eriksson L, Pyakurel P, Johansson O, Biberfeld P, Andersson S and
Isaguliants M. Autoimmunogenicity of a DNA-binding domain.
Mol Immunol 46, 1467, 2009.
Starodubova E, Boberg A, Ivanov A, Latyshev O, Petrakova N,
Kuzmenko Y, Litvina M, Chernousov A, Kochetkov S, Karpov V,
Wahren B and Isaguliants MG. Potent cross-reactive immune
response against the wild-type and drug-resistant forms of HIV
reverse transcriptase after the chimeric gene immunization.
Vaccine 28, 1975, 2010.
Masalova OV, Lesnova EI, Pichugin AV, Melnikova TM, Grabovetsky VV, Petrakova NV, Smirnova OA, Ivanov AV, Zaberezhny AD,
Ataullakhanov RI, Isaguliants MG and Kushch AA. The successful immune response against hepatitis C nonstructural protein
5A (NS5A) requires heterologous DNA/protein immunization.
Vaccine 28, 1987, 2010.
Department of Microbiology, Tumor and Cell Biology – MTC
HIV-1 and HIV-2 mediated patho­
genesis in relation to innate
immunity and immune activation
HIV-1 and HIV-2 are two
related viruses with distinct
clinical outcome. HIV-1 is
more pathogenic and transmissible than HIV-2. HIV-1
Marianne Jansson
is also spread worldwide
[email protected]
while HIV-2 largely remains
endemic in West Africa.
Disease progression of both infections is related to dysregulation of
the immune system. However, the mechanism behind the divergent
clinical outcomes is still to be revealed. In fact, the reason why HIV-2
in most cases is a controlled infection may give clues to how to modulate the pathogenesis of HIV-1. We have studied the impact of HIV-1
and HIV-2 on innate immunity function and chronic immune activation. Since microbial stimulation of Toll-like receptors (TLRs) triggers both anti-viral cytokines and initiate adaptive immunity we set
out to analyze TLR responsiveness.
We found that responsiveness to TLR7/8 stimuli is defective during
HIV-1 infection while maintained during HIV-2 infection. Responsiveness to other types of TLR stimuli was shown to be impaired with
loss of CD4+ T cells in both infections. Chronic immune activation
is a marker of disease progression and it was recently suggested that
immune activation in HIV-1-infected individuals results from microbial translocation into the blood after disruption of the gut mucosa.
Our studies showed that
microbial translocation,
assessed as lipopolysaccharide in plasma,
is independent of the
infectious agent, HIV-1
or HIV-2, but correlates
with severity of the disease.
To better understand
Microbial translocation correlates to severity of both HIV-1 and HIV-2 infections.
how HIV interacts with
innate immune cells for
efficient spread, we also studied the ability of HIV to utilize dendritic
cells (DCs), and the DC expressed molecule DC-SIGN, for transinfection of T cells. We found that HIV-1 variants emerging after
AIDS onset display reduced ability to use DC-SIGN, while direct T cell
infection efficacy is enhanced. This suggests that the ability of HIV-1
to use DC for efficient spread to T cells is more important during early
and chronic phases of infection than at end-stage disease.
Selected publications
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Borggren M, Repits J, Kuylenstierna C, Sterjovski J, Churchill MJ,
Purcell DF, Karlsson A, Albert J, Gorry PR and Jansson M. Evolution of DC-SIGN use revealed by fitness studies of R5 HIV-1 variants emerging during AIDS progression. Retrovirology 5, 28, 2008.
Nowroozalizadeh S, Månsson F, da Silva Z, Repits J, Dabo B, Pereira
C, Biague A, Albert J, Nielsen J, Aaby P, Fenyö EM, Norrgren H, Holmgren B and Jansson M. Studies on toll-like receptor stimuli responsiveness in HIV-1 and HIV-2 infections. Cytokine 46, 325, 2009.
Nowroozalizadeh S, Månsson F, da Silva Z, Repits J, Dabo
B, Pereira C, Biague A, Albert J, Nielsen J, Aaby P, Fenyö EM,
Norrgren H, Holmgren B and Jansson M. Microbial translocation
correlates to severity of both HIV-1 and HIV-2 infections. J Infect
Dis 201,1150, 2010.
Research Reports Infection Biology
Immune escape and the
potency of the cellular
immune system against HIV-1
The cellular immune system
includes T cells that can be
divided into two arms: CD8+
and CD4+. They battle virusinfected cells by recognizAnnika Karlsson
ing small peptides (epitopes)
[email protected]
of digested virus particles or
proteins displayed on the human
leucocyte antigen (HLA) alleles on an antigen presenting cell. The
interactions between the T cell and the HLA-peptide complex are
highly specific and thus extremely sensitive to variations. As the human
immunodeficiency virus (HIV) is highly variable this poses one of
the major obstacles in understanding the efficacy of a specific T cell
response. Thus, vaccine and treatment failures have been attributed to
the development of immune escape mutants and our inability to correlate viral control to pre-existing immune responses. A second obstacle
is the variation of the human immune system; a multitude of HLA alleles and T cell receptors (TCR) recognizing the HLA-peptide complex.
In my group, we study the interactions between the T cells and HIV
using immunological, virological, and bioinformatic approaches. We
hypothesize that a better understanding of the complex relationships
between the virus, host genetic factors, and the immune response will
provide valuable insights for the development of a vaccine.
For an effective antiviral response, we believe that a multitude of
viral epitopes would have to be matched by a multitude of human
HLA alleles to be effective. The recent advances in bioinformatics have
made it possible to evaluate some of the HLA-epitope-TCR interactions in greater details. Importantly, using this approach we have
identified a limited set of HLA class I epitopes eliciting broad population coverage as preferred by a vaccine antigen. We believe that this
knowledge can help us to identify regions to target in the next generation of HIV vaccines. Our goal is to identify epitopes within conserved
regions that are recognized by as many patients as possible and from
which the virus is unlikely to escape.
CD8+ T cells cripple HIV
replication by killing virus
infected cells and production of antiviral cytokines
(e.g. IFN-g, TNF-a). They
recognize, via their T
cell receptor (TCR), viral
epitopes presented on
the surface of HIV-infected cells by HLA class I
molecules.
Why do we get sick
when voles don’t?
Hantaviruses cause two severe zoonotic
diseases: hemorrhagic fever with renal
syndrome (HFRS) in Eurasia and hantavirus
cardiopulmonary
syndrome
Jonas Klingström
(HCPS) in the Americas. There are no
[email protected]
treatments or vaccines available, and
depending on the specific hantavirus up
to 40% of patients die during infection. Puumala virus, carried by
the bank vole, causes a relatively mild variant of HFRS, called nephropathia epidemica, in northern Europe including Sweden. There are
several other hantaviruses that also cause disease in man, all of them
have rodents as natural hosts. In rodents, infection causes a life-long,
but asymptomatic infection, while humans who are infected will get
the disease, but clear the infection.
It is not known why humans but not rodents get HFRS/HCPS.
HFRS/HCPS are believed to be caused by the immune response raised
against the virus,
rather than directly
by the virus infection
per se. By analyzing
immune responses in
patients and voles, we
hope to identify possible differences that
might explain why
we get disease while
rodents are protected.
We are also trying
Hantaan virus (green) infected cells show less
to characterize how
phosphorylated STAT1 (red) in nuclei (blue)
than non-infected cells after IFN-g-stimulation.
hantaviruses interfere
with the normal activation and function,
of innate and adaptive immune responses. We also want to investigate
the differences in the interference with the immune system in humans
and natural hosts.
We have found that hantaviruses are rather insensitive to the
antiviral action of interferons and human saliva. Hantaviruses also
avoid to activate innate immune responses against the infected cells.
A better understanding of the mechanisms behind the interference
of hantavirus with the human immune system and its disease causing properties is of importance for the development of HFRS/HCPSspecific treatments and vaccines.
Selected publications
Selected publications
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Karlsson AC, Chapman JM, Heiken BD, Hoh R, Kallas EG, Martin
JN, Hecht FM, Deeks SG and Nixon DF. Antiretroviral drug
t­herapy alters the profile of HIV-1-specific T cell responses and
shifts the immunodominant CTL response from Gag to Pol. J Virol
81, 11543, 2007.
Karlsson AC, Iversen AKN, Chapman JM, de Oliveira T, Spotts G,
McMichael AJ, Davenport M, Hecht FM and Nixon DF. Broadening CTL responses in early HIV-1 infection are associated with
viral escape. PLoS One 2, e225, 2007.
Pérez C, Larsen MV, Gustafsson R, Norström M, Atlas A, Nixon
D F, Nielsen M, Lund O and Karlsson AC. Broadly immunogenic
HLA class I supertype restricted elite CTL epitopes recognized
in a diverse population infected with different HIV-1 subtypes. J
Immunol 180, 5092, 2008.
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Stoltz M, Ahlm C, Lundkvist Å and Klingström J. Serum interferon
(IFN)-l is decreased in Hantavirus-infected patients and in vitro
established infection is insensitive to treatment with all IFNs and
inhibits IFN-g-induced nitric oxide production. J Virol 81, 8685,
2007.
Habjan M, Andersson I, Klingström J, Schümann M, Martin A,
Zimmermann P, Wagner V, Pichlmair A, Schneider U, Mühlberger
E, Mirazimi A and Weber F. Processing of genome 5’ termini as a
strategy of negative-strand RNA viruses to avoid RIG-I-dependent interferon induction. PLoS ONE 3, e2032, 2008.
Hardestam J, Lundkvist Å and Klingström J. Sensitivity of Andes
hantavirus to antiviral effect of human saliva. Emerg Infect Dis 15,
1140, 2009.
Department of Microbiology, Tumor and Cell Biology – MTC
47
Research Reports Infection Biology
Antimicrobial resistance
– new threats and
methodological progress
Modern health care depends on
the effective use of antimicrobials.
Increasing rates of antimicrobial
resistance noted all over the world
poses a serious threat to health care
Göran Kronvall
systems. We have a longstanding
[email protected]
interest in these problems focusing on analyzing resistance and on
methodological studies of susceptibility tests.
In an investigation of antimicrobial resistance in relation to socioeconomic factors in an area in Peru we found higher resistance in high
income groups compared to low income families. This was the most
interesting finding, but other results also deserved attention. In these
studies we used a modified rapid method to detect both resistance in
dominant Enterobacteriaceae bacteria in the gut as well as total resistance including resistant bacteria in low numbers (Kristiansson et al.,
2009).
In molecular studies of integron-carried resistance in Escherichia
coli we have discovered two new dfr-genes. Such genes give rise to
trimethoprim resistance. The appearance of these resistance genes
emphasizes the rapid development of new molecular types of resistance in a bacterial species, which is both a commensal in the gut and
also sometimes acts as a pathogen (Grape et al., 2007).
There has been a long series of studies in our laboratory of methods for antimicrobial susceptibility testing, focusing on the most
common test, the disc diffusion method. Single strain regression analysis (SRA) and the M-test are two analytical methods designed from
the results of these investigations. Normalized resistance interpretation (NRI) is the latest addition to this arsenal of analytical methods
for studying antimicrobial resistance. The method is now patented in
the USA (US Patent No. 7,465,559) and patents are pending in other
countries. Its use has recently been expanded into the area of marine
microbiology where lack of standards and scarcity of test results have
made interpretation of susceptibility difficult. NRI was the solution to
most of these problems (Smith et al., 2007).
Parasitic infections are
major obstacles to development
especially
in
Africa. Large-scale initiatives to control major
Ewert Linder
parasitic infections have
[email protected]
shown that misdiagnosis –
over-diagnosis in the case
of malaria and under-diagnosis of schistosomiasis - is a severe but
neglected problem. Basically this is due to poor resources: microscopy
in endemic regions is not cared for and standardization and quality
assurance of diagnostics are difficult to achieve.
To overcome these difficulties, we are developing microscopybased diagnostics, independent of ordinary microscopy. The aim is
to perform image capture of stool-, blood-, sputum- etc. specimens
at the grass root level and to carry out diagnostics by bi-directional
communication with a central server capable of computer vision,
image analysis and instant feedback.
As a first step towards these goals we have begun to explore the
possibilities of virtual microscopy as an educational and quality assurance tool for parasitology (Lundin et al., 2009). We have established a
website (http://www.webmicroscope.net/parasitology/), where parasite specimens for diagnostics can be viewed as in an ordinary microscope, and a network of European web servers has been developed to
ensure stabile high speed data transfer (Linder et al., 2008).
To develop virtual microscopy for
diagnostics of parasitic infections, we
aim at miniaturizing image capture
making the ordinary microscope obsolete. We have managed to obtain images
of helminth eggs with sufficient resolution for visual identification by placing
specimens on ordinary microscope
slides in contact with standard image
sensors (see Figure).
Our results suggest that helminth
On-chip imaging using
eggs such as those excreted in Schisstandard complementary
metal oxide semiconductosoma mansoni infection can be
tor (CMOS) image sensor
detected by on-chip imaging. Further
of digital camera (arrow),
technical developments are needed in
produces images of Schistosoma mansoni eggs in
order to obtain a resolution sufficient
stool specimen. Microscope
for the detection of smaller pathogens
slide with sample (a),
such as malaria parasites and specific
CMOS image sensor chip
(b), resulting digital image of
computer algorithms for image analythe sample (c and d). Schissis need to be worked out.
Selected publications
Selected publications
Escherichia
coli antimicrobial resistance against
gentamicin,
trimetoprim and
trimeroprimsulfamethxazole
between 2005 –
2008 at the Karolinska Hospital,
Solna.
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Direct image capture on sensor
chips and virtual microscopy for
diagnostics of parasitic infections
Smith P, Ruane NM, Douglas I, Carroll C, Kronvall G and Fleming
GTA. Impact of inter-lab variation on the estimation of epidemiological cut-off values for disc diffusion susceptibility test data for
Aeromonas salmonicida. Aquaculture 272, 168, 2007.
Grape M, Sundström L and Kronvall G. Two new dfr-genes in
trimethoprim-resistant integron-negative Escherichia coli. Antimicrob Agents Chemother 51, 1863, 2007.
Kristiansson C, Grape M, Gotuzzo E, Samalvides F, Chauca J, Larsson M, Bartoloni A, Pallecchi L, Kronvall G and Petzold M. Socioeconomic factors and antibiotic use in relation to antimicrobial
resistance in the Amazonian area of Peru. Scand J Infect Dis 41,
303, 2009.
Department of Microbiology, Tumor and Cell Biology – MTC
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tosome eggs sized about
50x100 micrometers (e).
Linder E, Lundin M, Thors C, Lebbad M, Winiecka-Krusnell J,
Helin H, Leiva B, Isola J and Lundin J. Web-based virtual microscopy for parasitology: a novel tool for education and quality assurance. PLoS Negl Trop Dis 2, e315, 2008.
Lundin M, Szymas J, Linder E, Beck H, de Wilde P, van Krieken H,
García Rojo M, Moreno I, Ariza A, Tuzlali S, Derviolu S, Helin H,
Lehto VP and Lundin J. A European network for virtual microscopy-design, implementation and evaluation of performance.
Virchows Arch 454, 421, 2009.
Derda M, Winiecka-Krusnell J, Linder MB and Linder E. Labeled
Trichoderma reesei cellulase as a marker for Acanthamoeba cyst wall
cellulose in infected tissues. Appl Environ Microbiol 75, 6827, 2009.
Research Reports Infection Biology
Research on
emerging viruses
Our research is based on an interdisciplinary approach between molecular virology,
immunology, genetics, epidemiology and
diagnostic aspects of zoonoses, especially
Åke Lundkvist
highly human-pathogenic viruses. Our
[email protected]
major efforts are at present focusing on the
following agents: Flaviviruses (tick-borne
encephalitis virus, dengue virus), Bunyaviruses (hantavirus, Rift
Valley fever virus) and avian influenza virus.
Our hantavirus program has generated important results concerning novel animal models (monkey and rodents), virus-host interactions, pathogenesis, apoptosis and innate immunity. We have also
found valuable, and sometimes surprising results on how, and under
which circumstances, various Bunyaviruses can survive outside their
vectors and hosts. The extreme increase of Epidemic Nephropathy
caused by Puumala hantavirus infections in northern Sweden during
recent years prompted us to investigate the presence and circulation
of viral genetic lineages/sublineages in more detail.
One of the major responsibilities of our group is microbiological
preparedness. Microarray-based diagnostic techniques for detection
of various emerging viruses have been developed. The awareness of
highly pathogenic avian influenza virus repeatedly infecting man
initiated also basic research aiming for a better understanding of the
transmission and the dramatic changes in virulence. Based on a new
animal model, we have studied how various avian influenza virus
strains affect one of the major reservoirs, the mallard.
Dengue fever pathogenesis and Rift Valley fever vaccine candidates are examples of new research areas. Based on a unique collection
of virus isolates from dengue fever, dengue hemorrhagic fever, and
dengue shock syndrome patients in Cambodia, we have recently initiated the search for markers hopefully explaining at least parts of the
dramatic variation in dengue pathogenesis. New approaches, based
on expression of viral structural proteins in plants, have been initiated
for Rift Valley fever vaccine development.
The Mallard
(”gräsand” in
Swedish) – an
important natural reservoir
of avian influenza virus.
Genetic variability
of important viral
pathogens
Viral hepatitis is a leading cause of liver
cancer and a common reason for liver
transplantation. We have previously
described three new genotypes E, F and
H of hepatitis B virus (HBV). MolecuLars Magnius
lar epidemiology of HBV worldwide
[email protected]
detected virus isolates with identical
S-genes common in Russia and Estonia.
These isolates remained closely related, when complete HBV genomes
were sequenced, but divided into five different clades three of which
shared a monophyletic origin pointing towards possible iatrogenic
transmission in the past (Tallo et al., 2008).
Dendrogram
obtained with
UPGMA based
on complete HBV
genomes showing evolutionary
relatedness and
monophyly of
three clades, I,
II and IV, with
identical S genes
within each clade
and the unrelated
clade V also with
identical S genes.
Hepatitis E virus (HEV) was once considered a pathogen confined
to tropical areas causing waterborn outbreaks often with thousands of
cases. Collaborative research has revealed indigenous cases in Sweden
and Denmark caused by HEV genotype 3. This genotype was prevalent not only in pigs, but also in wild boar which carried strains similar
to the human cases (Norder et al., 2009). The route of transmission
between pigs and man is under investigation.
Enteroviruses infect millions of individuals with a wide range of
clinical outcomes ranging from inapparent infections to acute flaccid paralysis. In parallel with the Center of Disease Control, USA,
we established for the first time molecular typing for enteroviruses
which yielded results consistent with serological typing enabling us
to identify new enterovirus types. In line with these studies we have
characterized enterovirus strains from cases of acute flaccid paralysis
suspected to be caused by the polioviruses, other enteroviruses, in
the Democratic Republic of Kongo. Thereby divergent isolates were
found representing two new enterovirus types, EV-93 and EV-94
(Junttila et al., 2007).
Selected publications
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Selected publications
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Hardestam J, Karlsson M, Falk KI, Olsson G, Klingström J and
Lundkvist A. Puumala hantavirus excretion kinetics in bank voles
(Myodes glareolus). Emerg Infect Dis 14, 1209, 2008.
Klingström J, Stoltz M, Hardestam J, Ahlm C and Lundkvist Å.
Passive immunization protects cynomolgus macaques against
Puumala hantavirus challenge. Antivir Ther 13, 125, 2008.
Jourdain E, Gunnarsson G, Wahlgren J, Latorre-Margalef N, Bröjer
C, Sahlin S, Svensson L, Waldenström J, Lundkvist Å and Olsen B.
Influenza virus in a natural host, the Mallard: Experimental infection data. PLoS ONE 5, e8935, 2010.
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Junttila N, Lévêque N, Kabue JP, Cartet G, Mushiya F, MuyembeTamfum JJ, Trompette A, Lina B, Magnius LO, Chomel JJ and
Norder H. New enteroviruses, EV-93 and EV-94, associated with
acute flaccid paralysis in the Democratic Republic of the Congo. J
Med Virol 79, 393, 2007.
Tallo T, Tefanova V, Priimägi L, Schmidt J, Katargina O, Michailov
M, Mukomolov S, Magnius L and Norder H. D2: major subgenotype of hepatitis B virus in Russia and the Baltic region. J Gen Virol
89, 1829, 2008.
Norder H, Sundqvist L, Magnusson L, Østergaard Breum S,
Löfdahl M, Larsen LE, Hjulsager CK, Magnius L, Böttiger BE
and Widén F. Endemic hepatitis E in two Nordic countries. Euro
Surveill 14, 19211, 2009.
Department of Microbiology, Tumor and Cell Biology – MTC
49
Research Reports Infection Biology
Molecular mechanisms
controlling adaptation
of bacteria
Bacteria need to sense the environment
in order to rapidly adapt to new and
changing conditions, for example when
they cause infections. We are especially
interested in how the membraneÖjar Melefors
bound two-component sensor BarA
[email protected]
can sense environmental factors and
how this leads to phenotypic changes
in the bacteria. It is not known exactly what these factors are, but
we have recently demonstrated that growth rate plays a clear role in
BarA activation in Escherichia coli. Activation of BarA leads to phosphorylation of the response regulator UvrY, which, in turn, controls
transcription of the CsrB and CsrC sRNAs. These two sRNAs act by
sequestering the CsrA regulatory protein. Free CsrA can bind to a
number of target mRNAs, leading commonly to a block in translation and a decreased mRNA stability (or in some cases to the opposite
effects). Earlier work has mainly identified target mRNAs involved in
carbon metabolism.
Using a microarray approach, we have recently shown that target
mRNAs in E. coli often encode GGDEF/EAL domain containing
proteins. We could, in a subsequent work, also show that CsrA targets
a different set of GGDEF/EAL mRNAs in Salmonella typhimurium.
Bacteria contain a large number of GGDEF and EAL domain proteins
that act by controlling the levels of the intracellular second messenger cyclic di-GMP. This second messenger seems to be a key factor in
the activation of virulence genes and in the decision of the bacteria to
remain in a biofilm state or to become motile. We believe that a better
understanding of these regulatory systems could reveal novel targets
for antibiotic therapy in the future.
Adaptation via the
BarA system in
Escherichia coli.
Selected publications
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Jonas K, Edwards AN, Simm R, Romeo T, Römling U and Melefors,
Ö. The RNA binding protein CsrA controls c-di-GMP metabolism by directly regulating the expression of GGDEF proteins. Mol
Microbiol 70, 236, 2008.
Jonas K and Melefors Ö. The Escherichia coli CsrB and CsrC
small RNAs are strongly induced during growth in nutrient poor
medium. FEMS Microbiol Lett 297, 80, 2009.
Jonas K, Edwards AN, Ahmad I, Romeo T, Römling U and Melefors Ö. Complex regulatory network encompassing the Csr, c-diGMP, and motility systems of Salmonella typhimurium. Environ
Microbiol 12, 524, 2010.
Department of Microbiology, Tumor and Cell Biology – MTC
Ecology of the intestinal
Escherichia coli flora
and diarrhoeal diseases
The aim is to increase the understanding of the ecology of the intestinal flora and the role of diarrhoeagenic Escherichia coli (DEC). We have
found that the majority of infant diarRoland Möllby
rhoea in León appears to be caused by
[email protected]
E. coli that contains various virulence
genes. There are different types of
DEC, the best studied are the enterotoxigenic E. coli (ETEC) that may
contain the gene encoding labile toxin (LT) and/or stable toxin (ST).
During diagnostic, the virulence genes are detected by PCR-based
methods using the collected sample, but not on individual isolates.
Strangely enough, though, the various DEC types were almost as
often detected in healthy infants as in infants with acute diarrhoea.
Even more, when we investigated the individual bacterial isolates
present in each individual by PhenePlate(R) typing, the strains diversity was similar in sick
and healthy infants.
Often only a small
fraction of the E. coli
bacteria investigated
were found to contain
the specific virulence
genes. In contrast, E.
coli strains containing the stable (entero)
toxin
gene
estA,
showed a low diversity, were only found Hypothesis of spread of virulence genes in true
in a limited number of or false DEC.
strains and dominated
the E. coli flora in children with diarrhoea. These strains were sometimes very similar to old ETEC reference strains, isolated in other
parts of the world more than twenty years ago. EstA carrying strains
were able to transfer their genes through conjugation to non-pathogenic E. coli strains in the test tube.
It is thus possible that there exist certain “Truly pathogenic E. coli
(TPEC?)” that are very stable over time and that are able to dominate the intestinal flora of E. coli when they cause diarrhoea. Furthermore, these strains are “promiscuous” and give away their virulence
genes in the intestine to various strains of the normal flora. These
“Falsely pathogenic E. coli” are then detected in faeces and erroneously interpreted as the cause of diarrhoea. Therefore, new and more
reliable methods for such investigations, from individual diagnostic
to national surveillance projects, need to be developed urgently.
Selected publications
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Reyes D, Vilchez S, Paniagua M, Colque P, Weintraub A, Möllby R
and Kühn I. Diversity of intestinal Escherichia coli populations in
Nicaraguan children with/without diarrhoea. J Med Microbiol 58,
1593, 2009.
Vilchez S, Reyes D, Paniagua M, Bucardo F, Möllby R and Weintraub A. Prevalence of diarrhoeagenic Escherichia coli in children
from León, Nicaragua. J Med Microbiol 58, 630, 2009.
Rahman M, Huys G, Kühn I, Rahman M and Möllby R. Prevalence
and transmission of antimicrobial resistance among Aeromonas
populations from a duckweed aquaculture based hospital sewage
water recycling system in Bangladesh. Antonie Van Leeuwenhoek
96, 313, 2009.
Research Reports Infection Biology
The normal
microflora
All healthy mammals are born germfree,
thus free of any microbes. Yet shortly after
birth, microbes establish themselves with
a growing complexity. Today we know that
Elisabeth Norin
the bacterial cells in the intestine outnumTore Midvedt
ber the number of cells comprising the
[email protected]
whole human body. Thus, the microbes
[email protected]
live in a symbiosis with the host, mostly in a
“balanced unbalance”, i.e. in a system based
on continuous cross talk. There are three major actors, namely the
host organism, the intestinal microflora and the environment. Any
alteration of these actors, such as antimicrobial treatment, various
diseases, surgery, physical-emotional stress, dietary alterations or
probiotics, malnutrition or starvation may disturb the balance for a
shorter or longer period.
The intestinal microflora can be evaluated in several ways. We have
chosen to study the flora under germfree and conventional conditions using the MAC-GAC concept. A MAC (Microflora Associated
Characteristic) is defined as any anatomical structure, physiological, immunological or biochemical function in an organism that has
been influenced by the microbes. When active microbes are absent,
as in healthy newborn infants, sometimes after antimicrobial treatment and in germfree animals, these functions are defined as GACs
(Germfree Animal Characteristics).
Today we are involved in several projects. We follow the establishment of the intestinal flora in infants, healthy and diseased, aiming
to investigate the role of exogenous factors of the development of
the flora, health status and eventual allergy development. We study
the intestinal flora in children at different ages from Italy, Germany,
Spain, Scotland and
Sweden. We are also
involved in investigations regarding
the role of the flora
in celiac disease.
Moreover, we investigate the role of
dietary supplements
(pre- and probiotics) given to both
infants and elderly.
Scheme of our gastrointestinal tract.
Selected publications
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Benno P, Ernberg I, Marcus C, Midtvedt T, Möllby R, Norin E and
Svenberg T. MAGEN. Bakterier, buller och brak. Karolinska Institutet University Press 2008.
Norin E, Jernberg C, Nilsson H-O and Engstrand L. Studies of
the intstinal microflora by traditional, functional and molecular
techniques. In; Lactobacillus, Molecular Biology – from genetics
to probiotics. Eds: Ljung Å, Wadström T. Caister Academic press
Norfolk, UK. p. 83, 2009.
Sandin A, Bråbäck L, Norin E and Björkstén B. Faecal short chain
fatty acid pattern and allergy in early childhood. Acta Paediatr 98,
823, 2009.
Microbe-host interactions
with a focus on Grampositive infections
Our main research focuses on pneumococcal infections that are major
causes of morbidity and mortality world-wide, annually killing
1–2 million individuals. PneumoBirgitta Henriques Normark
cocci are also common colonizStaffan Normark
ers of healthy children, a fact that
[email protected]
has prompted us to identify host
staffan [email protected]
and microbial factors deciding the
outcome of disease. We found capsular serotypes with a high invasive
disease potential behave as primary pathogens, whereas types with a
lower relative risk of causing invasive disease affect primarily patients
with underlying disease.
Comparative genomic analyses identified a large number of accessory genetic regions present in subsets of clinical isolates. One such
region codes for a pilus preferentially expressed by clones prone to
colonize the nasopharynx. This pilus contributes to virulence and
inflammatory host responses.
On the host side, Toll like receptor 9 plays a central role in early
pneumococcal defense promoting clearance by alveolar macrophages. Defensin-mediated neutrophil killing and bacterial capture
in neutrophil extracellular traps (NETs) have been studied in detail.
Pneumococci are not killed by NETs which are formed in pulmonary infections. NET trapping can be affected by the pneumococcal
capsule and the surface charge of the bacteria.
Furthermore, we have studied the role of antimicrobial effector
proteins and peptides in host defence of the lungs and small intestine
and described the first example of a complete developmental switch
in innate immune effector expression and anatomical distribution.
Moreover, we have
characterized pili in
enterococci, and studied Type III secretion
system inhibitors in
Chlamydia infections.
Finally, our recent data
suggest that cysteine
Pneumococci (green) are trapped in NETs
exclusion is an impor(elastase-red, DNA-blue).
tant adaptive strategy
among some Gram-positive bacteria against the challenges presented
by oxidative environments. The practical implications of our data
have facilitated antigen discovery for vaccines. Our studies on pilus
epidemiology, composition and formation provide a basis for a pilus
subunit vaccine and the structure/function of a pilus synthesizing
enzyme opens novel approaches to prevent pilus formation.
Selected publications
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Sjöström K, Blomberg C, Fernebro J, Dagerhamn J, Morfeldt E,
Barocchi M, Andersson M, Browall S, Henriques F, Rappuoli R,
Normark S and Henriques-Normark B. Clonal success of piliated
penicillin non-susceptible pneumococci. Proc. Natl Acad Sci USA
104, 12907, 2007.
Henriques-Normark B, Blomberg C, Dagerhamn J, Bättig P and
Normark S. The rise and fall of bacterial clones: with a focus on
Streptococcus pneumoniae. Nat Microbiol Rev 6, 827, 2008.
Daniels R, Mellroth P, Bernsel A, Neiers F, Normark S, von Heijne
G and Henriques-Normark B. Disulfide bond formation and
cysteine exclusion in Gram-positive bacteria. J Biol Chem 285,
3300, 2010.
Department of Microbiology, Tumor and Cell Biology – MTC
51
Research Reports Infection Biology
Characterization of HIV-1
populations in infected
cells: role in HIV-1 evolution
and persistent viremia
Determining the genetic relatedness of HIV-1 populations in cells,
plasma and tissue is important to
understand the pathogenesis of
HIV-1 and to target and eradicate
persistent HIV viremia in patients
Sarah Palmer
on suppressive therapy. We
[email protected]
adapted unique techniques never
used before in parallel, single-cell
sequencing, single-genome sequencing, and single-copy assay, to
characterize HIV-1 populations in single infected cells and plasma
from HIV-infected patients.
Analysis of cells and plasma from 9 HIV-infected patients revealed
that a majority (80-90%) of infected CD4+ T cells contained a single
viral DNA molecule indicating limited potential for recombination
in virus produced by these cells. Sequencing revealed that the intracellular viral DNA from CD4+ T cells was phylogenetically related to
contemporaneous plasma RNA implying ongoing exchange between
these compartments throughout infection. Moreover, the genetic
diversity and phylogenetic relatedness of intracellular and plasmaderived sequences from one patient remained unchanged after 6
months of suppressive therapy indicating a large pool of infected
cells persists even after plasma viral RNA decrease. Analysis of 20002x105 CD14+/CD16+ monocytes from the same patients revealed no
infected cells suggesting the frequency of infection is much greater in
CD4+ T cells than monocytes.
In collaboration with the US National Institutes of Health, we are
phylogenetically comparing the HIV populations in plasma to those
in infected cells extracted from peripheral blood mononuclear cells
and cells located in the lymphoid tissue. These studies will further
extend our understanding of HIV pathogenesis. With partners in
Spain, Germany, Netherlands and USA, we characterize persistent
HIV virus infection in patients on HIV treatment intensification to
determine if additional therapy reduces and possibly eradicates residual virus in plasma or other compartments such as the central nervous system.
Investigation of functionally
important antibodies
against malaria
Malaria is a disease that kills one
million people every year. Nearly
all deaths are caused by the parasite Plasmodium falciparum. The
merozoite form of P. falciparum
Kristina E M Persson
invades red blood cells and [email protected]
plies within the cell, finally causing rupture of the red blood cell
with release of daughter merozoites back into the circulation after 48
hours. Those who live in malaria-endemic areas and do not die from
the disease at a young age, eventually develop immunity, but only
slowly and after repeated exposure. Potential candidates for development of vaccines include antigens expressed on the surface of merozoites and/or proteins that are involved in erythrocyte invasion.
Antibodies are an important component of acquired protective
immunity. In our group, we are trying to understand which functions of antibodies are important for protection against malaria.
Basic understanding of how these antibodies work is imperative for
creating a functioning vaccine against malaria. We are studying the
affinity of antibodies directed against different merozoite antigens, as
well as the invasion inhibitory effect, and correlating this to protection. We are also investigating different invasion pathways in wild type
isolates of parasites, and correlating this to symptoms and severity of
disease. We have before developed a method for measuring invasion
inhibitory antibodies, and we have also shown that variation in use
of invasion pathways (involving the merozoite antigens EBA and Rh
proteins) mediates evasion of inhibitory antibodies.
The merozoite form of P. falciparum invades
erythrocytes, but this process can be inhibited by antibodies directed against different
merozoite antigens.
Similar diversity in
intra- and extracellular HIV compartments.
Selected publications
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Kearney M, Maldarelli F, Shao W, Margolick JB, Daar ES, Mellors
V, Rao V, Coffin JM and Palmer S. HIV-1 population genetics and
adaptation in newly infected individuals. J Virol 83, 2715, 2009.
Pereyra F, Palmer S, Miura T, Block BL, Wiegand A, Rothchild
AC, Baker B, Rosenberg R, Cutrell E, Seaman MS, Coffin JM and
Walker BD. Persistent low-level viremia in HIV-1 elite controllers
and relationship to immunologic parameters. J Infect Dis 200, 984,
2009.
Dahl V, Josefsson L and Palmer S. HIV reservoirs, latency, and reactivation: prospects for eradication. Antivir Res 85, 286, 2010.
Department of Microbiology, Tumor and Cell Biology – MTC
Selected publications
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Persson KE, McCallum FJ, Reiling L, Lister NA, Stubbs J, Cowman
AF, Marsh K and Beeson JG. Variation in use of erythrocyte invasion pathways by Plasmodium falciparum mediates evasion of
human inhibitory antibodies. J Clin Invest 118, 342, 2008.
Brolin KJ, Persson KE, Wahlgren M, Rogerson SJ and Chen Q.
Differential recognition of P. falciparum var2csa domains by
naturally acquired antibodies in pregnant women from a malaria
endemic area. PLoS One 5, e9230, 2010.
Persson KE. Erythrocyte invasion and functionally inhibitory
antibodies in Plasmodium falciparum malaria. Acta Trop 114, 138,
2010.
Research Reports Infection Biology
Inflammatory
responses to
bacterial pathogens
The ability of the host to defend itself
against invasion by respiratory bacterial pathogens depends on the capacity
to mount efficient immune responses.
Our research focuses on their role in the
Laura Plant
induction of innate immunity and in
[email protected]
priming of adaptive immune responses
to bacterial infection. We have recently
published studies on the effect of the pneumococcal cytotoxin, pneumolysin, on human dendritic cell (DC) responses.
Our studies revealed that the expression of haemolytic pneumolysin by pneumococci specifically inhibits the maturation of human
DCs and the induction of proinflammatory cytokines and activation
of the inflammasome. In addition, we have shown that clinical isolates
with non-hemolytic pneumolysin are more proinflammatory and
less apoptotic compared to clonally related strains with active pneumolysin. We have also been conducting studies onto the effect of the
pneumolysin and pneumococcal produced hydrogen peroxide on T
cell activation and survival.
Despite accumulating evidence that chronic inflammation associates with prostate cancer the responsible causes of prostatic inflammation remain elusive. Epidemiologically, prostatitis and prostate
cancer have been connected with infectious agents that evoke and
maintain persistent inflammation. In a collaborative project we are
evaluating the inflammatory potential of the prostate in relation to
bacterial infections and bacterial components in order to determine
whether bacterial infection can be implicated in events leading to
prostate cancer.
Intracellular localization of pneumococci in dendritic cells.
Selected publications
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Plant L, Wan H and Jonsson A-B. Non-lipooligosaccharide mediated signaling via toll-like receptor 4 causes fatal meningococcal
sepsis in a mouse model. Cell Microbiol 9, 657, 2007.
Lövkvist L, Sjölinder H, Wehelie R, Aro H, Norrby-Teglund A.
Plant L and Jonsson AB. CD46 contributes to the severity of group
A streptococcal infection. Infect Immun 76, 3951, 2008.
Littmann M, Albiger B, Normark S, Henriques-Normark B and
Plant L. Streptococcus pneumoniae evades human dendritic
surveillance by pneumolysin expression. EMBO Mol Med 1, 211,
2009.
Bacterial-host homeostasis
in the gastrointestinal tract
and oral cavity
Mammals have evolved in the
presence of bacteria. Hosts have
developed symbiotic relations
which, especially in the mouth
and gastrointestinal tract, allow
Mats Andersson
beneficial bacteria to reside, while
Katrin Pütsep
preventing opportunistic [email protected]
gens to gain foothold. The mechanism for this selection system is
largely unknown. Our hypothesis is that antimicrobial peptides are a
part of a system that controls this homeostasis.
The intestinal mucosal epithelium consists of a single cell layer.
This epithelium is rich in host derived antimicrobial peptides and
proteins, which kill bacteria without involvement of strong inflammatory, tissue damaging reactions. We have shown that the repertoire
and expression of antimicrobial peptides change following birth,
increases in the mouse small intestine from gut to colon (Karlsson,
Pütsep et al., 2008). The secreted intestinal antimicrobial components
are retained by the mucus layer (Meyer-Hoffert et al., 2008), thus indicating a role for these peptides in shaping the microflora. The clinical relevance is stressed by the fact that many patients with inflammatory bowels diseases present with low levels of intestinal peptides
and highly colonized mucus layers. To further elucidate the impact
of innate defence on the
natural microflora, we
will analyze the intestinal bacterial metagenome in gene knockout models for innate
defence molecules.
The protection of
the oral cavity is largely
mediated by neutrophils and individuals
with neutrophil dysfunction or severe neuImmunohistochemical localization of antibiotic peptides (CRS4C) in Paneth’s cells of
tropenia are at high risk
the mouse small intestine: Blue: cell nuclei.
for periodontal disGreen: CRS4C.
ease and tooth loss. We
have earlier demonstrated that neutrophil granules of patients with
chronic severe congenital neutropenia (SCN) are deficient in the antimicrobial peptide LL-37. This deficiency may serve as a specific biomarker for SCN. The lack of LL-37 was at a transcript level, although
expression was readily activated by vitamin D in neutrophil precursors (Karlsson, Carlsson et al., 2008). Further studies aim to elucidate
the mechanisms for this deficiency.
Selected publications
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Karlsson J, Pütsep K, Chu H, Kays RJ, Bevins CL and Andersson
M. Secreted enteric antimicrobial activity localizes to the mucus
surface layer. Gut 57, 764, 2008.
Karlsson J, Carlsson G, Larne O, Andersson M and Pütsep K. Vitamin D3 induces pro-LL-37 expression in myeloid precursors from
patients with severe congenital neutropenia. J Leukocyte Biol 84,
1279, 2008.
Meyer-Hoffert U, Hornef MW, Henriques-Normark B, Axelsson
L-G, Midtvedt T, Pütsep K and Andersson M. Regional variations
in Paneth cell antimicrobial peptide expression along the mouse
intestinal tract. BMC Immunol 17, 9, 2008.
Department of Microbiology, Tumor and Cell Biology – MTC
53
Research Reports Infection Biology
Microbial pathogenicity
– a functional integration of
virulence factor and bacterial
house-keeping functions
To enable colonization of hosts
bacterial pathogens rely on
selected genetic traits termed
virulence factors. Amongst
enteric bacteria, virulence factors
are often coded for by horizonMikael Rhen
tally acquired genetic elements in
[email protected]
the form of genetic continuums,
termed pathogenicity island that
became integrated into a conserved core genome during the evolution of the pathogen. Thus one may ask how such virulence factors
became functionally integrated into functions coded for by the core
genome of the bacterium. Others and we have shown that elemental
virulence factors do not act as independent units but strongly depend
on functions provided by the core genome. This integration occurs
at two levels. First, the genetic regulation of virulence genes has to be
integrated with the existing pattern of gene regulation. Second, the
synthesis and assembly of virulence factors also rely on the biochemical constitution of the bacterial cell.
We have used transcriptomic profiling of the classical bacterial
model pathogen – Salmonella enterica – to define how the expression
of individual genes on a whole genome scale becomes altered as the
pathogen shifts between host-like environments. Such analyses defined
significant alterations in virulence gene expression and revealed many
core genome genes to be co-regulated with virulence genes. Inactivation of such core genes may not affect the in vitro well-being of the
bacteria but may have a strong effect on the virulence in infection
models. An example of this is thioredoxin 1, an evolutionary highly
conserved oxidoreductase. Thioredoxin
is needed for the
activity of the Salmonella pathogenicity
island 2, which in Salmonella in its phase as an intracellular
bacterial pathogen. The image reveals S.
turn steers bacterial typhimurium bacteria (green) replicating in an
intracellular replica- eukaryotic cell with the aid of secreted virution and virulence. lence proteins (red). Host cell nuclei in blue.
Our intention is to
deepen such data mining and to probe for additional connections
between virulence genes and ordinary house-keeping genes. The findings will illuminate how pathogens have evolved to cause disease and
avoid host immune responses and possibly reveal new targets for pharmaceutical interference with virulence.
Selected publications
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Hautefort I, Thompson A, Eriksson-Ygberg S, Parker ML, Lucchini
S, Danino V, Bongaerts RJ, Ahmad N, Rhen M and Hinton JC.
During infection of epithelial cells Salmonella enterica serovar
Typhimurium undergoes a time-dependent transcriptional adaptation that results in simultaneous expression of three type 3 secretion systems. Cell Microbiol 10, 958, 2008.
Negrea A, Bjur E, Puiac S, Eriksson-Ygberg S, Åslund F and Rhen
M. Thioredoxin 1 participates in the activity of the Salmonella
enterica serovar Typhimurium pathogenicity island 2 type III
secretion system. J Bacteriol 191, 6918, 2009.
Puiac S, Negrea A, Richter-Dahlfors A, Plant, L and Rhen M.
Omeprazole antagonizes virulence and inflammation in Salmonella enterica-infected RAW264.7 cells. Antimicrob Agents Chemother 53, 2402, 2009.
Department of Microbiology, Tumor and Cell Biology – MTC
Cyclic di-GMP signaling in
biofilm formation and virulence
of Salmonella typhimurium
The small molecule cyclic
di-GMP is a novel secondary
messenger in many bacteria
that regulates biofilm formation and virulence. Regulation
Ute Römling
of cyclic di-GMP metabolism
[email protected]
is complex, for example the
genome of the enteric pathogen Salmonella typhimurium encodes eight putative di-guanylate
cyclases and fifteen putative phosphodiesterases.
Previously, we have shown that two di-guanylate cyclases contribute to the expression of biofilm formation in S. typhimurium. Recently,
we demonstrated that four phosphodiesterases play a determinative
role in the expression of biofilm formation. Thus, the effective cyclic
di-GMP concentration is determined by the delicate balance between
di-guanylate cyclase and phosphodiesterase activities.
Virulence is another phenotype frequently affected by cyclic
di-GMP signaling. We could recently show that cyclic di-GMP signaling affected invasion and induction of a pro-inflammatory cytokine
in epithelial cells, basic phenotypes that are major determinants of S.
typhimurium virulence. Invasion was mainly inhibited through the
biofilm regulator CsgD and the biofilm matrix component cellulose.
Inhibition of the pro-inflammatory response occurred through CsgD
which inhibited the secretion of pro-inflammatory monomeric flagellin.
The biofilm matrix component cellulose also affected microbehost interaction in the probiotic Escherichia coli strain Nissle 1917.
Cellulose production in Nissle 1917 contributes to enhanced cytokine
production and to adhesion of bacteria to a gastrointestinal epithelial
cell line and to the mouse epithelium in vivo.
Our fundamental research on bacterial signaling mechanisms
demonstrated that cyclic di-GMP signaling is not only required for the
regulation of biofilm formation, but also affected virulence properties
in S. typhimurium. Future research will focus on the molecular mechanisms of cyclic di-GMP signaling in biofilm-formation and virulence.
Salmonella typhimurium expressing cellulose.
Selected publications
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Simm R, Lusch A, Kader A, Andersson M and Römling U. The role
of EAL containing proteins in multicellular behavior of Salmonella enterica serovar Typhimurium. J Bacteriol 189, 3613, 2007.
Monteiro C, Saxena I, Wang X, Kader A, Bokranz W, Simm R, Nobles
D, Chromek M, Brauner A, Brown Jr RM and Römling U. Characterisation of cellulose production in Escherichia coli Nissle 1917 and
its biological consequences. Environ. Microbiol 11, 1105, 2009.
Lamprokostopoulou A, Monteiro C, Rhen M and Römling U.
C-di-GMP controls virulence properties of Salmonella enterica
serovar Typhimurium at the mucosal lining. Environ Microbiol
12, 40, 2010.
Research Reports Infection Biology
Molecular pathogenesis
of severe Plasmodium
falciparum malaria
We study the molecular pathogenesis of severe Plasmodium falciparum
malaria with a focus on the molecules
PfEMP1- RIFIN-SURFIN present at
the infected red cell surface. RIFINs
Mats Wahlgren
are co-transported with PfEMP1 and
[email protected]
SURFINs to the parasitized erythrocytes surface but also accumulate in
the parasitophorous vacuole and associate with the merozoite. By
exposing shared polymorphic antigens on both parasitized erythrocytes and merozoites the parasite may coordinate the antigenic
composition of these attachment-surfaces during growth in the
bloodstream. Indeed, a recent analysis predicted that the RIFINs have
undergone neo- or sub-functionalization and it was found possible
to divide the RIFINS into sub-groups with differential developmental expression and localization patterns. The multiple genes encoding
PfEMP1-RIFIN-SURFIN were identified to originate from duplications and transpositions of genomic segments onto multiple subtelomeres. The ubiquity of the segmental duplications, and the nature
of genes within, suggests an important role in plasmodia speciation.
Our team previously described P. falciparum rosetting, its association with severe malaria and the importance of the PfEMP1-family
of proteins as adhesins. Rosetting depends on PfEMP1 and heparan
sulfate expressed at the erythrocyte- and endothelial cell surfaces.
Based on these findings we are presently developing a receptor-based
drug together with Dilafor AB, a KIAB-company (www.dilafor.com).
A significant milestone was recently passed with the completion of a
Phase I study proving that the new compound is safe and well tolerated. Furthermore, an archetype anti-rosetting vaccine against severe
disease based on the heparan sulfate-binding domain of PfEMP1
is presently being developed. Immunization of animals abolished
PfEMP1-mediated sequestration. These results support the use of
the PfEMP1 domain in the development of a vaccine targeting severe
malaria.
Staining of two
distinct SURFIN
in the parasitophorous vacuole
of Plasmodium
falciparum (red
and green).
Nuclei are stained
blue.
Towards protective
and curative HIV
immunity
We are working on strategies to prevent
or treat HIV infection by immunological means. For this purpose we have
constructed and refined genetic vaccines
representing several subtypes of HIV as
Britta Wahren
well as including most genes of the virus.
[email protected]
Compositions of genes are inserted in
single plasmids. When a suitable composition, according to subtypes resident in the geographical region, is
introduced into animal cells, they produce both free proteins and
virus-like particles. Preclinical studies of the properties of these
vaccine plasmids have demonstrated broad and strong cell-mediated
and humoral reactivity systemically and at mucosal tissues. We have
shown protection to HIV challenge in an experimental model, and
even been able to eradicate
infectious HIV by means of
HIV-directed drug-labeled
antibody infusion. Our clinical HIVIS program has set
out to study the delivery of
this genetic HIV vaccine
boosted with a recombinant
vaccinia vector in Sweden
and Tanzania. Strong and
broad cross reactivity against
antigens from HIV subtypes
HIV virus-like particle recovered
A-E have been found as well
from Vero cells transfected with DNA
as good safety. Although the
plasmids encoding HIV genes gp120,
gp41, and gag. Anti V3 P4/D10+ staph
HIV Vaccine ImmunogenicA labelled gold particle attaches to
ity Study vaccine schedule
surface of HIV-like particle (upper right
was designed to elicit primaquadrant). Photomicrograph by B.
Wahren, D. Hallengärd, A. Bråve, M.
rily cell-mediated immunity,
Liu, L. Xing and H. Cheng.
broad antibody responses
are observed in all volunteers after boosting. Therapeutic studies in Sweden and Italy have
revealed induction of new cell-mediated immune responses as well
as strengthening of previous reactions. Our next goal is to amplify
primary responses by improved delivery systems, both systemically
and locally at the mucosae of humans.
Selected publications
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Selected publications
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Normark J, Nilsson D, Ribacke U, Winter G, Moll K, Wheelock CE,
Bayarugaba J, Kironde F, Egwang TG, Chen Q, Andersson B and
Wahlgren M. PfEMP1 motifs predict severity of Plasmodium falciparum malaria. Proc Natl Acad Sci USA 104, 15835, 2007.
Chêne A, Donati D, Guerreiro-Cacais AO, Levitsky V, Chen Q, Falk
KI, Orem J, Kironde F, Wahlgren M and Bejarano MT. A molecular link between malaria and Epstein-Barr virus reactivation. PLoS
Pathog 3, e80, 2007.
Mok BW, Ribacke U, Rasti N, Kironde F, Chen Q, Nilsson P and
Wahlgren M. Default pathway of var2csa switching and translational repression in Plasmodium falciparum. PLoS ONE 3, e1982,
2008.
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Bråve A, Boberg A, Gudmundsdotter L, Rollman E, Hallermalm K,
Ljungberg K, Blomberg P, Stout R, Paulie S, Sandström E, Biberfeld
G, Earl P, Moss B, Cox JH and Wahren B. A new multi-clade DNA
prime/recombinant MVA boost vaccine induces broad and high
levels of HIV-1-specific CD8(+) T cell and humoral responses in
mice. Mol Ther 9, 1724, 2007.
Sandström E, Nilsson C, Hejdeman B, Bråve A, Bratt G, Robb
M, Cox J, Vancott T, Marovich M, Stout R, Aboud S, Bakari M,
Pallangyo K, Ljungberg K, Moss B, Earl P, Michael N, Birx D,
Mhalu F, Wahren B, Biberfeld G and the HIV Immunogenicity
Study 01/02 Team. Broad immunogenicity of a multigene, multiclade HIV-1 DNA vaccine boosted with heterologous HIV-1
recombinant modified vaccinia virus Ankara. J Infect Dis 198,
1482, 2008.
Roos AK, Eriksson F, Timmons JA, Gerhardt J, Nyman U,
Gudmundsdotter L, Bråve A, Wahren B and Pisa P. Skin electroporation: effects on transgene expression, DNA persistence and local
tissue environment. PloS One 4, e7226, 2009.
Department of Microbiology, Tumor and Cell Biology – MTC
55
Research Reports Cell and Tumor Biology
Cell and Tumor Biology Research
Hanahan and
Weinberg published the highly influential
and visionary millennium article on the
“Hallmarks of Cancer”, where 25 years of
cancer research was summarized (Hanahan D and Weinberg RA, Cell 100, 57, 2000).
Cancer was described as a multistep process
involving activating/inactivating mutations
in oncogenes and tumor suppressor genes
(TSG), respectively, resulting in deregulation
of fundamental cellular processes, thereby
leading to cancer. Common hallmarks of
cancer cells of different origins included
Exactly ten years ago
56
self-sufficiency in growth signals, insensitivity to anti-proliferative signals,
evading apoptosis, limitless replicative
potential, sustained angiogenesis and
tissue invasion and metastasis (see
Figure). Despite tremendous progress
to elucidate these processes, cancer
metastasis is still poorly understood
and will surely come more into focus.
Ten years later, a number of new
hallmarks can be added to the list as a
result of the accelerating scientific and
technological development (see Figure). One
Marie Arsenian
Henriksson
Pontus
Aspenström
Yihai Cao
Ingemar Ernberg
Georg Klein
Sonia Laín
Lars-Gunnar
Larsson
Uno Lindberg
Sven Pettersson
Galina Selivanova
Laszlo Szekely
Department of Microbiology, Tumor and Cell Biology – MTC
Eugene
Zabarovsky
such hallmark is block of differentiation,
which characterizes most tumors. This may
be related to the discovery of cancer stem cells
or tumor-initiating cells with self-renewal
capacity within the tumor mass. Another
important hallmark of tumor cells which is
under intense research is their potential of
epigenetic reprogramming of the genome,
which may explain some of their plasticity.
A remarkable recent finding was that only
Eva Klein
four genes, including two oncogenes, could
induce reprogramming of differentiated cells
into pluripotent stem cells (iPS), an epigenetic phenomenon that may have relevance
for cancer. Another hallmark is the ability of
tumor cells to evade oncogene-induced cellular senescence, a state of irreversible growth
arrest. This is probably related to another hallmark, the ability to cope with cellular stress
causing DNA damage responses, one of the
features of “oncogene addiction”. It has also
become increasingly clear during recent years
Rolf Ohlsson
that communication between tumor cells and
the environment, such as the tumor stroma
Project leaders:
is imperative to sustain growth and facilitate
Gesan Arulampalam metastasis. Avoiding immune surveillance is
LiFu Hu
yet another hallmark.
Stefan Imreh
Much of the recent achievements
Elena Kashuba
mentioned above have been made possible
through the very rapid technological development in genomics as well as in other “omics”
enabling high-throughput measurements
at a global level. This has paved the way for
“systems biology”, where all these data are integrated, facilitating identification of key regula-
Research Reports Cell and Tumor Biology
tors that will be in focus as targets for cancer
drug development. The development of RNAi
technology and the expanding field of microRNA discovery greatly facilitate this work.
From basic to translational research
Laín, Marie Arsenian Henriksson, Ingemar
Ernberg, Georg Klein, Elena Kashuba and
Lars-Gunnar Larsson. Pontus Aspenström
is studying the family of Rho GTPases and
their role in cell signaling, as well as in cell
and organelle movements. Uno Lindberg is
performing basic research on the action of the
microfilament system of the cell.
Our tumor and cell biology research connects
to several of the areas mentioned above and
represents both research on fundamental
biological questions to translational research.
EBV, p53 and Myc in focus
Rolf Ohlsson’s group performs world leading
Since the days at the Department of Tumor
epigenetic investigations on the organization
Biology, EBV has been used as a tool to underof the nucleus and higher order chromatin
stand oncogenic transformation of cells in
structure. The groups of Eugene Zabarovsky
the studies of Eva Klein, Georg Klein, Ingeand LiFu Hu are also carrying out epigemar Ernberg, Elena Kashuba, Laszlo Szekely
netic
characterizaand LiFu Hu. Significant
tions of tumor cells.
work in the area of angi“Enhancing the collaboraStefan Imreh, Eugene
ogenesis in relation to
tions between all the cell and
Zabarovsky and Georg
cancer and metastasis as
tumor biology researchers
Klein perform detailed
well as to other diseases
with all our diverse skills
studies on identificais carried out by the
and expertise will give MTC
tion of new tumor
group of Yihai Cao. Sven
a very strong international
suppressor genes on
Petterson and Gesan
platform for successful cancer
chromosome 3. FuncArulampalam are studyresearch also in the future.”
tional studies on tumor
ing gut microbial signalsuppressor proteins and
ing in the regulation of
oncoproteins such as p53, Myc and oncogenes
inflammation and metabolism and its influencoded by the Epstein-Barr virus (EBV) and
ence on cancer, obesity and other diseases.
their role in tumorigenesis are performed
Strong efforts to identify and characterize
by the groups of Galina Selivanova, Sonia
small molecular compounds that reactivate
or enhance the function of p53 or that inhibit
the oncogenic and/or enhance the anti-tumorigenic functions of Myc is carried out by the
groups of Galina Selivanova and Sonia Laín
(p53) and Marie Arsenian Henriksson and
Lars-Gunnar Larsson (Myc) through cellular and/or interaction screens. The pioneering work of Galina Selivanova in this field
has brought the p53 reactivating molecule
PRIMA to clinical trials. Laszlo Szekely is
performing ground breaking combinatorial
drug screens based on new advanced imaging
technology.
KICancer Network
Last but not least, MTC scientists are interacting with other cancer researchers at KI
through the KICancer Network that organizes several seminars and yearly meetings.
Ingemar Ernberg is the enthusiastic driving
force for these meetings and in the seminar
serie on “What is life?” which has such an
important influence on the intellectual environment and interactions at MTC and KI as a
whole. Enhancing the collaborations between
the cell and tumor biology researchers with
all our diverse skills and expertise will give
MTC a very strong international platform for
successful cancer research also in the future.
Lars-Gunnar Larsson
Department of Microbiology, Tumor and Cell Biology – MTC
57
Research Reports Cell and Tumor Biology
Gut microbial signaling
in inflammation and
metabolism
The human gut harbors a large
number of different commensal
microbes, now recognized as
determinants of growth, nutritional, immune and metabolic
Velmurugesan Arulampalam
thresholds of the host. Many of
[email protected]
these physiological responses
are interdependent and are
linked to intestinal health. The use of germfree animals, housed and
maintained by our group, has facilitated the study of gut bacterial
impact on host biology.
The crosstalk between inflammation and metabolism has been
apparent for nearly a century. The nuclear receptor Peroxisome
Proliferator Activated Receptor g (PPARγ) is one of the most studied nuclear receptors in the context of inflammation and metabolism.
This receptor plays a central role in mediating adipogenesis and regulating glucose homeostasis.
More recently our laboratory observed that certain commensal bacterial strains can modulate the function of this receptor to
promote a robust anti-inflammatory response. The multi-faceted
properties of PPARg
renders it an ideal candidate to study how
gut microbial input
can regulate metabolic
thresholds.
We are now studying the crosstalk
between the gut epithelium and motile
immune cells in the
lamina propria, which
provide one natural
conduit for bacterial Can the balance between pro-inflammatory and
anti-inflammatory signals emanating from the
signals from the gut gut flora impact on cells of the lamina propria,
to different organs. e.g. macrophages and dendritic cells, which
We have postulated may determine the outcome of inflammationlinked metabolic lesions such as diabetes and
that functions in other atherosclerosis? Is it possible to tilt this balorgans may be affected ance in favour of a healthier outcome, through
by these microbial sig- manipulation of the gut microflora?
nals and metabolites.
The mechanisms behind these signals are enigmatic, and are thus relevant questions to be addressed, in relation to xenobiotic metabolism,
inflammation, diabetes and even cognitive functions.
Approaches to target
the MYC oncoprotein to
combat cancer
Neuroblastoma (NB), a tumor that
arises from neural crest cells in the
developing sympathetic nervous
system, is the most common and
deadly solid childhood tumor outside
Marie Arsenian Henriksson
the CNS. NB cells show complex
[email protected]
patterns of genetic defects, including
specific chromosomal aberrations and
MYCN amplification. This occurs in 40-50% of high-risk cases and is
associated with advanced-stage disease, rapid tumor progression and
a survival rate of less than 15%. An alternative treatment option for
these children is therefore urgently needed.
Our research is focused on the MYCN oncogene in order to identify targets for development of novel anti-cancer therapies. To this end
we will: 1) analyze the functional significance of MYCN-regulated
microRNAs for NB pathogenesis; 2) characterize the role of MYCN in
invasion and migration; 3) identify small compounds that selectively
kill cells with high with high MYCN expression; 4) explore the normal
function of MYCN during development of the nervous system.
It was recently shown that MYCN regulates expression of microRNAs in NB cells. We have demonstrated that miR-18a and miR19a from the oncogenic miR-17~92 cluster target and subsequently
repress the expression of estrogen receptor α (ESR1). Downregulation of mir-18a as well as ectopic ESR1 expression resulted in
growth arrest and neuronal differentiation of NB cells. Importantly,
we demonstrated expression of ESR1
in human fetal sympathetic ganglia
suggesting a role during neuronal development. We propose that MYCN amplification may disrupt estrogen-signaling
sensitivity in primitive sympathetic
cells through downregulation of ESR1,
thereby preventing normal neuroblast
differentiation.
Collectively, our studies will give miR-18a expression detected
with Cy3 (red) at embryonic
insights to the normal function of day E6 in a transverse secMYCN and its contribution to NB tion of the trunk region of the
pathogenesis. This knowledge could developing chicken embryo.
The image shows intense
serve as basis for development of novel miR-18 staining in the nervcancer therapies for children with neu- ous system (neural tube and
roblastoma as well as other diseases, dorsal root ganglia) and in the
dermomyotome. Cell nuclei
since MYC is activated in many different were counterstained with
DAPI (blue).
malignancies.
Selected publications
Selected publications
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Are A, Aronsson L, Wang S, Greicius G, Lee YK, Gustafsson JA,
Pettersson S and Arulampalam V. Enterococcus faecalis from
newborn babies regulate endogenous PPARγ activity and IL-10
levels in colonic epithelial cells. Proc Natl Acad Sci USA 105, 1943,
2008.
Lundin A, Bok CM, Aronsson L, Björkholm B, Gustafsson JA, Pott
S, Arulampalam V, Hibberd M, Rafter J and Pettersson S. Gut flora,
Toll-like receptors and nuclear receptors: a tripartite communication that tunes innate immunity in large intestine. Cell Microbiol
10, 1093, 2008.
Arulampalam V. Gastrointestinal inflammation: lessons from
metabolic modulators. J Intern Med 263, 607, 2008.
Department of Microbiology, Tumor and Cell Biology – MTC
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Albihn A, Mo H, Yang Y and Henriksson M. Camptothecininduced apoptosis is enhanced by Myc and involves PKCδ signaling. Int J Cancer 121, 1821, 2007.
Hydbring P, Bahram F, Su Y, Tronnersjö S, Högstrand K, von der
Lehr N, Lilischkis R, Hein N, Wu S, Vervoorts J, Henriksson M,
Grandien A, Lüscher B and Larsson LG. Myc/Ras cooperativity in
transformation: Phosphorylation by cdk2 is required for Myc to
repress Ras-induced senescence. Proc Natl Acad Sci USA 107, 58,
2010.
Lovén J, Zinin N, Wahlström T, Müller I, Brodin P, Fredlund E,
Ribacke U, Pivarcsi A, Påhlman S and Henriksson M. MYCNregulated microRNAs repress estrogen receptor α (ESR1) expression and neural differentiation in human neuroblastoma. Proc
Natl Acad Sci USA 107, 1553, 2010.
Research Reports Cell and Tumor Biology
Signaling networks
controlled by small
GTPases
The Rho GTPases are key regulators
of cell morphogenesis and cell migration. Current cladistic analysis has
shown that the family consists of 20
members, which can be further divided
Pontus Aspenström
into two classes, the classical and the
[email protected]
atypical Rho GTPases. The atypical Rho
GTPases function in a distinct manner
from the classical Rho GTPases since they reside constitutively in the
active, GTP-bound, conformation, which has profound signaling
consequences.
We want to elucidate the signaling pathway downstream of Rho
GTPases and Miro GTPases that control cell growth and cell migration during normal physiological conditions as well as during disease.
We have employed the yeast two-hybrid system to identify novel binding partners for all members of the Rho GTPases. This way, we identified the non-receptor tyrosine kinase Pyk2 as a binding partner for the
atypical Rho member Wrch-1. Our data show that Pyk2 collaborates
with Src family kinases in order to impose the Wrch-1-dependent
cellular effects.
The Miro GTPases, which we identified a couple of years ago, have
emerged as essential regulators of mitochondrial morphogenesis and
motility along microtubules. The Miro GTPases function as calciumdependent sensors in the control of mitochondrial motility. We have
identified novel binding partners to the Miro GTPases and we aim to
study their roles in model organisms, such as zebrafish. Several lines
of indications suggest that Miro GTPases are involved in neurodegenerative diseases. We intend to investigate if Rho GTPases and Miro
GTPases can function as targets for medical treatments for diseases
such as cancer and neurological pathologies.
Miro-dependent aggregation of mitochondria.
Transiently transfected
Miro1 is localized to mitochondria in COS7 cells
and does not influence
the morphology of the
mitochondrial network. In
contrast, transient transfection of a constitutively
active Miro1 mutant (MiroV13) result in mitochondrial aggregation caused
by the deregulation of
mitochondrial trafficking
and fusion/fission.
Angiogenesis in
cancer and nonmalignant diseases
Malignant and non-malignant human
disorders share a common mechanism
of switching on an angiogenic phenotype
in pathological settings. Thus, antiangiogenic drugs can be used for the treatment
Yihai Cao
of cancer and non-malignant diseases. In
[email protected]
fact, antiangiogenic drugs have become
an important component of the first-line
therapy against age-related macular degeneration and a variety of
human cancers. With this background, our research activity can be
summarized as follows:
1. Cancer and metastasis. We have
found that the growth factors FGF-2
and PDGF-B could synergistically
induce angiogenesis, tumor growth
and metastasis. We have demonstrated that growth factor VEGF
could stimulate tumor cell dissemination, invasion, and hematologous
and lymphatic metastasis. Impor- Blood vessel in brown adipose
tantly, tumor-derived angiogenic tissue.
factors could induce a systemic
destructive effect on multiple tissues and organs, manifesting cancer
cachexia, loss of body mass, and paraneoplastic, cancer-associated
disease, syndromes, which significantly jeopardize the quality of life
and shorten life time. We are one of the first to propose ”off-tumor”
targets as one of the mechanisms of antiangiogenic drugs.
2. Angiogenesis and obesity. We are at the leading position to
study the role of angiogenesis in adipogenesis and obesity. Using
both genetic and epigenetic animal models, we have identified crucial
angiogenic pathways leading to neovascularization in both white and
brown adipose tissues (WAT and BAT). A recent example is the coldinduced angiogenesis during transition from WAT into BAT.
3. Cardiovascular diseases. We continue to work on the molecular mechanisms underlying arteriogenesis in ischemic muscles. We
are the first to propose combinations of angiogenic and arteriogenic
factors for the treatment of ischemic myocardium after heart infarction.
4. Retinopathy. We have developed an adult zebrafish retinopathy
model, which allows us to study mechanisms of retinal angiogenesis
under physiological and low oxygen concentrations. This model also
allows us to identify new drugs for therapy.
Selected publications
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Selected publications
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Ruusala A and Aspenström P. The atypical Rho GTPase Wrch1
collaborates with the non-receptor tyrosine kinases Pyk2 and Src
in regulating cytoskeletal dynamics. Mol Cell Biol 28, 1802, 2008.
Ruusala A, Pawson T, Heldin C-H and Aspenström P. Nck is
involved in formation of dorsal ruffles, cell migration and cell
adhesion downstream of the PDGF b receptor. J Biol Chem 283,
30034, 2008.
Reis K, Fransson Å and Aspenström P. The Miro GTPases: at the
heart of the mitochondrial transport machinery. FEBS Letters 583,
1391, 2009.
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Nissen LJ, Cao R, Hedlund E-M, Wang Z, Zhao X, Wetterskog
D, Funa K, Bråkenhielm E and Cao Y. FGF-2 triggers PDGF-Binduced angiogenesis and their reciprocal interplay synergistically
promotes tumor neovascularization and metastasis. J Clin Invest
117, 2766, 2007.
Xue Y, Religa P, Cao R, Hansen AJ, Lucchini, Jones B, Wu Y, Zhu
Z, Pytowski B, Liang Y, Zhong W, Vezzoni P, Rozell B and Cao Y.
Anti-VEGF agents significantly confer survival advantages of
tumor-bearing mice by improving a cancer-associated systemic
syndrome. Proc Natl Acad Sci USA 105, 18513, 2008.
Xue Y. Petrovic N, Cao R, Larsson O, Chen S, Feldman HM, Liang
Z, Zhu Z, Nedergaard J, Cannon B and Cao Y. Hypoxia-independent cold-induced angiogenesis in brown and white adipose tissues:
mutually opposing roles of VEGF receptor-1 and -2 in modulating
adipose metabolism. Cell Metabolism 9, 99, 2009.
Department of Microbiology, Tumor and Cell Biology – MTC
59
Research Reports Cell and Tumor Biology
From viral molecular
switches to human
cancer
The interest of my group is viral infections associated with cancer risk in
man. Our prime system of study is the
Epstein-Barr virus (EBV) infection.
EBV infection is the most common
Ingemar Ernberg virus infection in humans world-wide.
[email protected]
Under conditions of immune dysregulation, which can be due to infections
with other pathogens, due to immunosuppression or genetic predisposition the EBV virus gets involved in carcinogenesis. About 200,000
new cases/year worldwide are estimated to primarily originate from
an EBV virus infection. The EBV virus can cause different types of
cancers with hematopeiotic or epithelial cell origin. We study primarily two molecular switches excerted by viral proteins that contribute
to cancer risk. One is the switch between cell proliferation and rest in
virus infected cells. The other switch is the switch between migration
of epithelial cells induced by the viral protein LMP2a in interaction
with the cellular tyrosine kinase Syk and their non-migration.
We apply our studies at the molecular level on EBV-associated
cancers: Hodgkins lymphoma, post-bone-marrow transplant
patients and HIV-carriers with high lymphoma risk and nasopharyngeal carcinoma.
We also use the EBV-infection of cells for a systems biology
approach to cancer. The hypothesis is formulated as the “Cancer Cell
Attractor” (se Huang et al., 2009).
Functional studies on human
chromosome 3 to understand
tumor progression
Virtually all tumors contain numerical and/or structural chromosome aberrations. During
the microevolution of a malignant cell clone the selection for
Stefan Imreh certain aberrant chromosome
[email protected] constitution leads to the loss of
certain chromosome regions
that may contain tumor growth inhibitory genes or tumor suppressor genes or to the gain or amplification of other regions that may
contain tumor growth promoting genes or oncogenes. We focus on
the functional relevance of chromosome 3 aberrations in various
malignancies. For this purpose we had previously developed a model
system called “elimination test”, based on chromosome 3 transfer into
tumor cells by microcell fusion. The microcell hybrids were tested for
the elimination versus retention of specific chromosome regions after passages in severe combined immunedeficiency (SCID) mice. We
identified a common eliminated region on chromosome 3 designated
CER1 that was similar in both human chromosome 3/mouse fibrosarcoma and chromosome 3/human RCC microcell hybrids. It contains 33 genes whereby 6 of them were discovered and cloned by our
group. Several of these genes may act as tumor suppressors. The group
focuses on the functional analysis of 3 out of potentially 7 tumor suppressor genes in CER1: LF, LIMD1 and TMEM7.
We are characterizing the CER1 and other chromosome 3 breakpoints. These breakpoint regions are “evolutionarily plastic” i.e. they
are involved in the evolutionary rearrangements of the chromosomes,
show intense transposon recruitment and contain segmental and/
or gene duplications. The puzzle of “evolutionary sorting” of tumor
suppressor versus tumor breakpoint regions should be solved as a
next step.
Using EBV-infected cell lines to evaluate the Cancer Attractor Hypothesis.
Selected publications
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Zhang X, Sanmun D, Hu L, Fadeel B and Ernberg I. Epstein-Barr
virus-encoded LMP1 promotes cisplatin-induced caspase activation through JNK and NF-κB signaling pathways. Biochem
Biophys Res Commun 360, 263, 2007.
Birgersdotter A, Baumforth KR, Porwit A, Sjöberg J, Wei W,
Björkholm M, Murray PG and Ernberg I. Inflammation and tissue
repair markers distinguish the nodular sclerosis and mixed cellularity subtypes of classical Hodgkin’s lymphoma. Br J Cancer 101,
1393, 2009.
Huang S, Ernberg I and Kauffman S. Cancer attractors: a systems
view of tumors from a gene network dynamics and developmental
perspective. Semin Cell Dev Biol 20, 869, 2009.
Department of Microbiology, Tumor and Cell Biology – MTC
Giemsa stained chromosomes.
Selected publications
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Kost-Alimova M and Imreh S. Modeling non-random deletions in
cancer. Semin Cancer Biol 17, 19, 2007.
Kost-Alimova M, Darai-Ramqvist E, Yau WL, Sandlund A, Fedorova L, Yang Y, Kholodnyuk I, Cheng Y, Li Lung M, Stanbridge E,
Klein G and Imreh S. Mandatory chromosomal segment balance
in aneuploid tumor cells. BMC Cancer 7, 21, 2007.
Darai-Ramqvist E, Sandlund A, Müller S, Klein G, Imreh S and
Kost-Alimova M. Segmental duplications and evolutionary plasticity at tumor chromosome break-prone regions. Genome Res 18,
370, 2008.
Research Reports Cell and Tumor Biology
Epigenetic changes
in initiation and
development of cancers
Epigenetics refers to the study of
heritable changes in gene expression
without a change in DNA sequence.
Epigenetic events are critical for
normal development and growth of
LiFu Hu
cells. In cancer, silencing of tumor
[email protected]
suppressor genes (TSG) or activation
of oncogene are main mechanisms
for carcinogenesis. The change in gene expression is often caused by
aberrant DNA methylation of CpG islands and histone hypoacetylation in early stages of cancer development, affecting various fundamental pathways, such as apoptosis, invasion and metastasis.
We found that promoter hypermethylation of the cellular genes
RASFF2 and CDH13 which work as TSGs is linked to clinical outcome
of nasopharyngeal carcinoma (NPC) as a tumor model. On the other
hand, hypomethylation activates the oncogene BAGE. Cancer related
Epstein-Barr virus also uses epigenetic mechanisms to maintain virus
latency and regulates the expression of both viral and cellular genes by
the EBV encoded LMP1 oncogene.
Aiming at an early diagnosis of cancer, we have identified novel
TSGs by methylation and expression microarrays, and developed a
sensitive “Multiplex Methylation Specific PCR (MMSP)” for NPC on
mouth washing/swab samples. MMSP was patented in Australia 2008
and is in clinical trail. Based on the same strategy we designed tests
to detect prostate/bladder cancer from urine and lung cancers from
sputum samples.
Micro RNAs (miRNAs) are short non-coding RNAs, which regulate gene expression in a sequence-specific manner via translation
inhibition or messenger RNA degradation, which contributes cancer
development
and
progression. Function of miRNAs in
tumor progression,
regulation by epigenetic alteration, and
potential application
for diagnosis of NPC
are being explored.
The EBV encoded,
170 bp long noncoding RNA EBERs,
which are expressed
abundantly
and
constitutively in the
Epigenetic changes and disease.
nucleus of all latently
infected cells, are also
studied.
EBNA-binding cellular
proteins and their role
in cell transformation
The project is aimed on elucidation of
the role of cellular proteins that bind
to Epstein-Barr virus (EBV)-encoded
nuclear antigens (EBNAs) in cell transformation.
Elena Kashuba
We are using the EBV-induced B cell
[email protected]
immortalization as a model system
to study the cell transformation. It is
clear now that EBV exploits the normal signaling pathways of the
B lymphocyte. The six growth transformation associated EBNAs
promote cell proliferation and protect from apoptosis. Studying the
influence of some EBNAs on the pRb – E2F1 pathway, we have shown
that EBNA-6 interacts with a pRb-associated protein, MRPS18-2
(S18-2), leading to the liberation of E2F1. This stimulates the entry of
the cell into the S-phase.
Unexpectedly, overexpression of S18-2 protein in the primary
rat embryonic fibroblasts (REFs) led to REF immortalization. The
immortalized cells (18IM) lose contact inhibition, form foci, and are
capable of anchorage-independent growth. Concurrently, mesodermal markers, such as vimentin, smooth muscle actin, and Fut4, disappear completely. 18IM cells express embryonic stem cell markers,
such as SSEA-1, Sox2, and Oct3/4. In immunodeficient mice, 18IM
cells formed small transiently growing tumors that have down-regulated SSEA-1 and showed pan-keratin staining.
Over the last few years, we have discovered 15 previously unknown
interactions between three transforming virus proteins, EBNA-3,
EBNA-5 and EBNA-6 and cellular host proteins. We have used the
yeast two-hybrid system, GST-pull down assay, immunoprecipitation with the following masspectrometry, and surface plasmon
resonance (SPR) to find
EBNA binding cellular
partners and to monitor
protein-protein binding.
To clarify the mechanism
of S18-2 induced immortalization we plan to use
knock-out cells (p53,
pRb etc) for transformation assay and the Danio
rerio model for S18-2
silencing and overexpression. Also we are studying
Immunostainings on primary rat embryonic
the properties of other
fibroblasts (REFs), c-Myc and Ras transproteins of S18 family,
formed cells (MR) and S18-2 transformed
cells (18IM) for mesenchymal (SMA,
S18-1 and S18-3.
vimentin) and stem (SSEA1) cell markers.
Selected publications
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Selected publications
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Sun D, Zhang Z, Van Do N, Ernberg I, Huang G and Hu L.
­Methylation of CDH13 promoter in NPC could serve as a potential diagnositic biomarker. Oral Oncol 43, 82, 2007.
Hu LF, Qiu QH, Fu SM, Magnusson K, Sun D, He B, Lindblom A
and Ernberg I. A region on chromosome 5q carries a susceptibility
locus predisposing for NPC. Eur J Hum Genet 16, 343, 2008.
Nguyen-Van D, Ernberg I, Phan-Thi Phi P, Tran-Thi C and Hu L.
Epstein-Barr virus genetic variation in Vietnamese patients with
nasopharyngeal carcinoma: full-length analysis of LMP1. Virus
Genes 37, 273, 2008.
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Kashuba E, Yurchenko M, Yenamandra SP, Snopok B, Isaguliants
M, Szekely L and Klein G. EBV-encoded EBNA-6 binds and targets
MRS18-2 to the nucleus, resulting in the disruption of pRb-E2F1
complexes. Proc Natl Acad Sci USA 105, 5489, 2008.
Savchenko A, Yurchenko M, Snopok B and Kashuba E. Study of
the spatial architecture of p53, MDM2, and p14ARF-containing
protein complexes. Mol Biotechnol 41, 270, 2009.
Kashuba E, Yenamandra SP, Darekar SD, Yurchenko M, Kashuba V,
Klein G and Szekely L. MRPS18-2 protein immortalizes primary
rat embryonic fibroblasts and endows them with stem cell like
properties. Proc Natl Acad Sci USA 106, 19866, 2009.
Department of Microbiology, Tumor and Cell Biology – MTC
61
Research Reports Cell and Tumor Biology
Role of EpsteinBarr virus in
lymphomas
Epstein-Barr virus (EBV) was discovered in
Burkitt lymphoma (BL) 50 years ago. The virus
carried by almost all humans is the causative
agent of infectious mononucleosis, IM. Nine
virally encoded proteins are expressed in B
Eva Klein lymphocytes and transform, in concert with
[email protected] cellular proteins, these cells into immortal cell
lines in vitro. In vivo they are recognized and
eliminated by the immune response. Under conditions of immunosuppression they may grow progressively.
Several tumors that originate in other cell types were also found to
carry the viral genome, but they do not express the full set of proliferation driving EBV proteins. The viral proteins that are expressed can
induce phenotypic changes in the cell and influence its interactions
with the microenvironment. We have discovered that soluble factors
produced in the environment can promote the development of malignancies such as Hodgkins and NK lymphomas by modifying the expression of virally encoded proteins and/or by promoting cell growth.
The response of humans to primary EBV infection is variable. It
can pass without any disease or it can induce IM. Our in vitro experiments suggest that the symptomatology of IM reflects the activation
of innate immunity. Leukotriene B4, known mainly for its role in
allergy, contributes to the activation of innate immunity.
We have also studied the hereditary EBV specific immunodeficiency X-linked lymphoproliferation (XLP). Boys affected with
XLP carry a mutated SAP with the lack of the proapopototic function. The lack of SAP function contributes to the understanding of
the clinical picture as activation of induced cell death is important for
lymphocyte homeostasis.
BL is driven by accidental chromosomal immunoglobulin/c-myc
translocation and exists in both EBV carrying and EBV negative forms.
Constitutive activation of myc that drives the cell to proliferate and
induces apopotosis. As SAP is only expressed in EBV carrying but not
in EBV negative
BL SAP promoted
apoptosis may be
counteracted
by
the anti-apoptotic
effect of EBNA-1
in the EBV genome
carrying cells which
would thus determine the fate of the
Hypothetical scheme of the contribution of SAP
Ig/myc transloca- and EBV to the fate of germinal center B cells with
tion carrying B cells. Ig/c-myc translocation.
Selected publications
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Liu A, Claesson H-E, Mahshid Y, Klein G and Klein E. Leukotriene B4 activates T cells which inhibit B cell proliferation in EBV
infected cord blood derived mononuclear cell cultures. Blood 111,
2693, 2008.
Nagy N, Matskova L, Kis LL, Hellman U, Klein G and Klein E. The
proapoptotic function of SAP provides a clue to the clinical picture
of X-linked lymphoproliferative disease. Proc Natl Acad Sci USA
106, 11966, 2009.
Kis LL, Persson EK, Salamon D, Nagy N, Scheeren FA, Spits H,
Klein G and Klein E. IL-21 imposes a type II EBV gene expression
on type III and type I B cells by the repression of C- and activation
of LMP-1-promoter. Proc Natl Acad Sci USA 107, 872, 2010.
Department of Microbiology, Tumor and Cell Biology – MTC
Epstein-Barr virus, oncogenes
and tumor suppressor genes
and immune and non-immune
surveillance against cancer
Georg Klein
[email protected]
The group works on the following questions: i) To analyze the
transforming effect of EpsteinBarr virus (EBV) for human
lymphocytes; ii) To study the
tumor suppressor regions of
human chromosome 3p; iii)
To identify contact mediated
microenvironmental control
against tumor development.
In the EBV field, we have identified, with Elena Kashuba, 10 new
interactions between EBNA-3, EBNA-5, and EBNA-6 and cellular
proteins. In addition, we have discovered that the cellular protein
MRPS18-2 can transform normal embryonic fibroblasts into
immortalized stem cell-like cells. We have shown, with Barbro EhlinHenriksson, that EBV influences chemokine receptors or chemokines
and that these modulate the migration of lymphoblasts. In the search
for tumor suppressor genes, we have identified, with Stefan Imreh
and Eugene Zabarovsky, ”asymmetric” tumor suppressor genes (our
term) that can inhibit tumor growth in vivo, without impairing the
growth of the same cells in vitro. These genes are of great interest, since
they may provide a lead for further investigations on the microenvironmental control of tumor growth. In a third area, I have defined
four different forms of non-immune surveillance: genetic (DNA
repair), epigenetic (stringency of chromatin structure), intracellular
(apoptosis, elicited by illegitimately activated oncogenes) and intercellular (microenvironmental control). With Laszlo Szekely we have developed a high throughput in vitro system that permits the detection of
normal fibroblast mediated inhibition of tumor cell growth.
Plans: In the EBV project, we are looking for additional interactions of EBNAs with cellular proteins. In the tumor suppressor field,
we wish to identify further ”asymmetric suppressors”. In the surveillance field, we wish to analyze the differences between the inhibitory
action of normal cells taken from the different body sites, from people
of different ages and different cancer risks, against prostatic carcinoma cell lines.
GFP-S18-2 (green signal) shows predominantly cytoplasmic distribution.
Upon EBNA-6 expression (blue signal, lower panel) GFP-S18-2 is translocated to the nucleus where it co-localizes with EBNA-6.
Selected publications
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Kashuba E, Yurchenko M, Yenamandra SP, Snopok B, Isaguliants
M, Szekely L and Klein G. EBV-encoded EBNA-6 binds and targets
MRS18-2 to the nucleus, resulting in the disruption of pRb-E2F1
complexes. Proc Natl Acad Sci USA 105, 5489, 2008.
Klein G. Toward a genetics of cancer resistance. Proc Natl Acad Sci
USA 106, 859, 2009.
Ehlin-Henriksson B, Liang W, Cagigi A, Mowafi F, Klein G and
Nilsson A. Changes in chemokines and chemokine receptor
expression on tonsillar B cells upon Epstein-Barr virus infection.
Immunology 127, 549, 2009.
Research Reports Cell and Tumor Biology
Discovery, characterization
and optimization of bioactive
small molecules through a p53
activation assay
Our group is interested in finding new small molecule therapeutics as well as chemical
tools for basic research using
Forward Chemical Genetics,
i.e. cell-based screens. So far
Sonia Laín
we have screened over 34,000
[email protected]
compounds for their ability
to increase the activity of the
p53 tumor suppressor in cells using a robust and inexpensive primary
assay followed by a carefully chosen series of secondary tests. p53 activation was chosen as a primary assay for two main reasons. First, p53
senses a wide variety of stress types and therefore, its activation can
be used to identify compounds that affect a variety of cell signaling
pathways involved in cancer as well as in other diseases. Second, there
are numerous high quality reagents to study p53, which improves
our chances of elucidating the mechanism(s) by which a compound
works in cells.
Several hits from our screening effort were selected for chemical
optimization through or collaboration with Nick Westwood’s lab in
Scotland. To date we have shown that two improved derivatives from
our initial set of hits decrease tumor growth rate in vivo. One is a potent tubulin depolymerizing agent called JJ78:12. The other, named
tenovin-6 (see figure), is the main focus of our current work. In order
to elucidate the mechanism of action of tenovin-6 we performed a
yeast genetic screen, biochemical assays and target validation studies
in mammalian cells. In this way we showed that tenovin-6 inhibits
the protein-deacetylase activities of SirT1 and SirT2, two members
of the sirtuin family of NAD+ dependent deacetylases linking metabolism and gene expression. We are currently performing studies
to further assess the selectivity of tenovin-6 and its derivatives for
sirtuins in cells, to
identify their binding site in the sirtuins, and to design
derivatives that are
more potent and/
Structure of tenovin-6.
or specific for each
of the sirtuin family
members. In addition, we have recently started a new screen using an
improved version of our p53 activation primary cell-based assay.
Selected publications
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Staples OD, Hollick JJ, Campbell J, Higgins M, McCarthy AR,
Appleyard V, Murray KE, Baker L, Thompson A, Ronseaux S,
Slawin AM, Lane DP, Westwood NJ and Laín S. Characterization,
chemical optimization and anti-tumor activity of a tubulin poison
identified by a p53-based phenotypic screen. Cell Cycle 7, 3417,
2008.
Laín S, Hollick JJ, Campbell J, Staples OD, Higgins M, Aoubala
M, McCarthy A, Appleyard V, Murray KE, Baker L, Thompson A,
Mathers J, Holland SJ, Stark MJ, Pass G, Woods J, Lane DP and
Westwood NJ. Discovery, in vivo activity, and mechanism of action
of a small-molecule p53 activator. Cancer Cell 13, 454, 2008.
Medda F, Russell RJ, Higgins M, McCarthy AR, Campbell J, Slawin
AM, Lane DP, Laín S and Westwood NJ. Novel cambinol analogs
as sirtuin inhibitors: synthesis, biological evaluation, and rationalization of activity. J Med Chem 52, 2673, 2009.
The c-Myc oncoprotein:
Function, regulation
and targeting
The myc oncogenes encode powerful
transcription factors that control expression of a vast number of genes involved in, among other functions, cell
growth, apoptosis, metabolism and
Lars-Gunnar Larsson
stem cell function. Deregulated MYC
[email protected]
contributes to the development of a
high proportion of human cancers, in
particular at the advanced stages of disease. We are studying the function and regulation of the Myc proteins, with the aim of identifying
molecules targeting Myc protein activity. Recently, we uncovered
regulation of cellular senescence, one of the main barriers of tumor
development, as a new function of Myc. We found that induction
of a Myc antagonist caused cellular senescence in Myc-transformed
cells (Wu et al., 2009). Further, Myc represses senescence induced by
other oncogenes such as Ras.
Repression of senescence by
Myc requires phosphorylation of Myc at Ser-62 by the
cell cycle kinase Cdk2 (Hydbring et al., 2010). Cdk2 is
also required to represses senescence induced by overexpressed Myc itself (Campaner et al., 2010). Importantly,
inhibition of Cdk2 by small
Bimolecular fluorescence complemenmolecule inhibitors induced
tation (BiFC) assay showing interaction
senescence in Myc-driven
between c-Myc and the E3 ubiquitin
ligase Skp2 in the nucleus of living cells
tumor cells (Hydbring et al.,
2010; Campaner et al., 2010).
Our future research aims
to elucidate exactly how Myc and Cdk2 regulates senescence and to
evaluate the in vivo potential of Cdk2 inhibitors in Myc-driven mouse
tumor models. Another potential way of combating Myc is to explore
and utilize mechanisms of Myc protein destruction via the ubiquitin/
proteasome pathway, and we are continuing our work to identify new
E3 ubiquitin ligases targeting Myc. Our recent research also involves
identification of low molecular weight inhibitors of interactions
between Myc and cofactors crucial for Myc function. Using novel
methods for visualizing protein interactions in living or fixed cells,
we have screened chemical libraries and identified several molecules
selectively targeting Myc. The long-term aim of this work is the development of new drugs for cancer treatment.
Selected publications
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Wu S, Hultquist A, Hydbring P, Cetinkaya C, Öberg F and Larsson
L-G. TGF-β enforces senescence in Myc-transformed hematopoietic tumor cells through induction of Mad1 and repression of Myc
activity. Exp Cell Res 315, 3099, 2009.
Campaner S, Doni M, Hydbring P, Verrecchia A, Bianchi L,
Sardella D, Schleker T, Perna D, Tronnersjö S, Murga M, Fernandez-Capetillo O, Barbacid M, Larsson L-G and Amati B. Cdk2
suppresses cellular senescence induced by the myc oncogene. Nat
Cell Biol 12, 54, 2010.
Hydbring P, Bahram F, Su Y, Tronnersjö S, Högstrand K, von der
Lehr N, Lilischkis R, Hein N, Wu S, Vervoorts J, Henriksson M,
Grandien A, Lüscher B and Larsson L-G. Phosphorylation by
Cdk2 is required for Myc to repress Ras-induced senescence in
cotransformation. Proc Natl Acad Sci USA107, 58, 2010.
Department of Microbiology, Tumor and Cell Biology – MTC
63
Research Reports Cell and Tumor Biology
Studies on the action of
the microfilament system:
Redox regulation of actin
and the tropomyosins
We study the structure and
function of the actin microfilament (MF)-system. Lately, our
focus has been on two aspects of
its dynamic regulation in nonmuscle cells: namely processes
Uno Lindberg
related to changes in the redox
[email protected]
status of actin, and mechanisms
coupled to the function of tropomyosin. We have found that redox control of actin by H2O2 can reversibly regulate polymerization and depolymerization of actin filaments.
H2O2 in nanomolar concentrations is sufficient to activate cysteines
of cytoplasmic actin and profilin for subsequent glutathionylation,
which likely have drastic consequences for their cellular function.
We have shown that a fraction of tropomyosin appears as soluble
isoform-specific multimers seemingly involved in actin reorganization after cell stimulation. Tropomyosin can access and control the
(+)-end of growing actin filaments where actin monomers bound
to profilin are added to drive cell edge advancement. It appears that
profilin and tropomyosin cooperate in fine-tuning this process which
yet remains to be elucidated at the molecular level. Interesting in
this context are the isoforms of tropomyosin where the variants of
the protein that localize to advancing lamellipodia differ from those
found along contractile actomyosin bundles further in from the edge.
Of particular concern is the extensive alteration of expression of
these isoforms in conjunction to malignancy. This supports the view
of a close connection between the control of the MF-system and the
altered behaviour of cancer cells, and suggests that alterations in the
expression of the tropomyosin isoform pattern is part of this connection.
Rat mammary
adenocarcinoma
cells (MTLn3)
expressing variants
of tropomyosin 5 (red
and green), nuclei
displayed in blue.
Photo: Li-Sophie
Rathje
Selected publications
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Lassing I, Schmitzberger F, Björnstedt M, Holmgren A, Nordlund
P, Schutt CE and Lindberg U. Molecular and structural basis for
redox regulation of β-actin. J Mol Biol 370, 331, 2007.
Lindberg U, Karlsson R, Lassing I, Schutt CE and Höglund AS. The
microfilament system and malignancy. Semin Cancer Biol 18, 2,
2008.
Grenklo S, Hillberg L, Zhao Rathje LS, Pinaev G, Schutt CE and
Lindberg U. Tropomyosin assembly intermediates in the control of
microfilament system turnover. Eur J Cell Biol 87, 905, 2008.
Three-dimensional
epigenetics: The
cancer connection
The eukaryotic cell nucleus holds keys to
many secrets of life. For example, the DNA
blueprint is condensed more than 100,000fold from its linear shape to the very limited
physical confines of the nuclear structure.
Rolf Ohlsson Despite this tight packaging, the higher
[email protected]
order chromatin structure must allow
regulated access to selected parts of the
genome to enable specific transcription profiles during developmental windows. There must also be a substantial measure of precision
in these processes, as chromatin features must be faithfully copied to
re-enact conformations pivotal for the memory of a cell lineage, and
to enable progressive restrictions in cell fates during development.
The epigenetic reprogramming process governs the fate of the developing mammalian
zygote by regulating
the early allocation
of the three germ
layers and many
other later pivotal
processes during
the lifetime of the
individual. Upon
abnormal environmental cues, such
as viral infection, The image shows the close physical proximities
between CTCF, a regulator of chromosomal netepigenetic instabil- works and replication timing patterns, with PARP1,
ity may follow. This which is associated with DNA repair. We believe
condition may both that the activity of PARP1, which is stimulated by
CTCF, indirectly influences the ability of CTCF to
initiate cancer and regulate replication-timing patterns (see also Farenable the selec- rar et al., Mol Cell Biol 30, 1199, 2010.)
tion of meta-stable
epigenetic marks sustaining abnormal proliferation at ectopic sites
during the malignant process.
It is becoming increasingly clear that the epigenome influences
the expressivity of the genome in 3 dimensions within the nuclear
architecture. Using novel techniques, the group is exploring reprogramming of chromosomal networks in human embryonic stem cells
and in relationship with other epigenetic features, such as replication
timing, replication origins and epigenetic marks of the primary chromatin fibres. We have thus been able to document that one replication
origin, the H19 imprinting control region regulates not only genome
wide patterns of interactions in a manner reflecting epigenetic reprogramming, but also the replication process of the sequences it interacts with. The overall aim of these projects is to understand the 3rd
dimension of the epigenome and how it relates to both normal development and cancer.
Selected publications
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Department of Microbiology, Tumor and Cell Biology – MTC
Göndör A and Ohlsson R. Replication timing and epigenetic
reprogramming of gene expression: a two-way relationship? Nat
Rev Genet 10, 269, 2009.
Göndör A and Ohlsson R. Chromosome crosstalk in three dimensions. Nature 461, 212, 2009.
Sandhu KS, Shi C, Sjölinder M, Zhao Z, Göndör A, Liu L, Tiwari
VK, Guibert S, Emilsson S, Imreh MP and Ohlsson R. Non-allelic
transvection of multiple imprinted loci is organized by the H19
imprinting control region during germline development. Genes
Dev 23, 2598, 2009.
Research Reports Cell and Tumor Biology
Gut microbiota: A key
regulator of body function
that sustains health
Bacteria have a symbiotic relationship with their host and this interplay is involved in the setting of an
organism’s homeostasis throughout life. Genetic variations and
Sven Pettersson
environmental cues that disrupt
[email protected]
this homeostasis are of paramount
importance in influencing health.
We have established an experimental platform including various
animal models (also germ free mice) and access to clinical material to
try to understand the imapct of gut micorbiota in health and disease.
Simple questions like: How does bacteria connect and communicate
with the host and which signaling pathways are used? Our hypothesis is that nuclear receptors (NR) act as second messengers in the
communication between the host and its microbiota. NR, in addition,
regulate and fine-tune most of the organs in the body.
Identification of biomarkers connected to homeostasis. In collaboration with the Genome Institute of Singapore, we are exploring the
impact of the incoming microbiota on host physiology in order to
identify host biomarkers that are regulated by the resident flora in various organs in the body. Additional projects aim to identify biomarkers and mechanisms underlying colorectal cancer and inflammatory
bowel disease (IBD).
The Core Facility for Germ Free Research (CFGR). I am also the director of the KI sponsored core facility for germ-free research. Rodents
are raised in an environment without gut microbiota and today we
have developed state of the art technology to derive new lines on
demand. The facility works in close collaboration with a number of
domestic and international academic partners including the biotech
industry.
A working model of gut microbiota and effects on normal developmental programming. Environmental cues are known to impact
the composition of the flora. The resultant altered flora may therefore be a risk factor in genetic predisposed inviduals.
Pharmaceutical
reactivation of p53 as a
strategy to combat cancer
p53 reinstatement leads to impressive regression of established
tumors in mice, supporting the
idea that restoring p53 is a good
strategy in cancer treatment. My
Galina Selivanova research focuses on the [email protected]
ment of small molecules restoring the tumor suppression functions of p53, either by refolding mutant p53 to rescue its activity,
or via preventing proteasomal degradation of p53 in tumors with
non-mutated p53. One of our molecules, PRIMA-1MET, which can
rescue the tumor suppressor function of mutant p53, is currently
being tested in first-in-man Phase I clinical trial. We are addressing
the fundamental question that need to be solved for the development
of novel medicines, i.e. understanding of the mechanism of action of
the candidate compounds, including target specificity in vitro and in
vivo and possible off-target effects.
Further, using small molecules as research tools, we address important questions of p53 biology. Applying systems biology we discovered important mechanisms which control the p53 choice between
induction of apoptosis and growth
arrest. We show that
MDM2-dependant
degradation of p53
cofactor
hnRNP
K and cdk inhibitor p21 switches
Ablation of key oncogenic pathways by RITAthe p53 response reactivated p53. We found a potent inhibition of
towards cell death crucial oncogenes by p53 upon reactivation by
(Enge et al., 2009). small molecule RITA using microarray analysis
(left). Inhibition of oncogenes by p53 reduces the
Further, we found cell’s ability to buffer pro-apoptotic signals and
that upon phar- elicits robust apoptosis (right).
macological activation, p53 unleashes a transcriptional repression of anti-apoptotic
proteins Mcl-1, Bcl-2, MAP4, and survivin, blocks the Akt pathway,
which is central in the transmission of growth regulatory signals
originating from cell surface receptors and c-myc oncogene on several
levels and downregulates the oncogene product cyclin E and the transcription factor β-catenin (Grinkevich et al., 2009). Our study adds
a new dimension to p53 regulation of physiological events, demonstrating that p53 reactivation triggers ablation of crucial oncogenes.
The multitude of oncogenes inhibited by p53 and multiple levels on
which they are targeted create external robustness of the p53 response
(see Figure for more details).
Selected publications
Selected publications
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Björkholm B, Bok CM, Lundin A, Rafter J, Hibberd ML and
Pettersson S. Intestinal microbiota regulate xenobiotic metabolism in the liver. PloS One 4, e6958, 2009.
Genander M, Halford MM, Xu NJ, Eriksson M, Yu Z, Qiu Z,
Martling A, Greicius G, Thakar S, Catchpole T, Chumley MJ,
Zdunek S, Wang C, Holm T, Goff SP, Pettersson S, Pestell RG,
Henkemeyer M and Frisén J. Dissociation of EphB2 signaling pathways mediating progenitor cell proliferation and tumor
suppression. Cell 139, 679, 2009.
Dermot PB et al., 2009. Genome-wide association identifies 28
ulcerative colitis susceptibility loci. Nat Genet 42, 332, 2010.
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Enge M, Bao W, Hedström E, Jackson SP, Moumen A and
Selivanova G. MDM2-dependent downregulation of p21 and
hnRNPK provides a switch between apoptosis and growth arrest
induced by pharmacologically reactivated p53. Cancer Cell 15,
171, 2009.
Grinkevich VV, Nikulenkov F, Shi Y, Enge M, Bao W, Maljukova A,
Gluch A, Kel A, Sangfelt O and Selivanova G. Ablation of key oncogenic pathways by RITA-reactivated p53 is required for efficient
apoptosis. Cancer Cell 15, 441, 2009.
Zhao CY, Szekely L, Bao W and Selivanova G. Rescue of p53 Function by small-molecule RITA in cervical carcinoma by blocking
E6-mediated degradation. Cancer Res 70, 3372, 2010.
Department of Microbiology, Tumor and Cell Biology – MTC
65
Research Reports Cell and Tumor Biology
High throughput live
cell imaging in cancer
research and therapy
The greatest paradox of cancer
therapy is that it has to be evidence
based and individualized at the same
time. In vitro chemosensitivity assays
have been suggested to resolve this
Laszlo Szekely
paradox. However, early assays were
[email protected]
not able to keep the tumor cells alive
for sufficiently long time. We have
developed a new cell culture system (OmniSanguine) that is based on
human whole blood extract that permits three days survival and a fluorescence multi-parameter functional analysis of primary tumor cells.
We have built and programmed automated fluorescence microscopes
(Hexascope) that can analyze cells on 384-well cell culture plates.
Our assay can test the sensitivity for thirty different drugs on a robot
printed drug plate on single cell level. We receive live tumor samples
from several European oncological/hematological centers. We have
initiated both retrospective and prospective studies to analyze different hematological malignancies (acute myeloid leukemias, acute and
chronic lymphoid leukemias, post transplant lymphomas, multiple
myelomas, late stage Hodgkin disease) as well as late stage carcinomas
that grow in the form of ascites or pleural effusion. Our ambition is
to provide drug sensitivity data for individualized assay guided therapies.
We also use this technology to identify tumor types that could
be targeted by newly developed drugs, as well as to screen chemical
libraries on live tumor cells to identify novel anti-cancer agents.
In addition we use color labeling of living cells – by chemical (fluorescent dyes) or genetic (transfection with fluorescent protein coding
genes) means – to study the interaction of tumor cells with immune
effector cells or normal stromal cells in mixed cell culture system in
high throughput assays.
Automated
measurement
of viability of
primary human
tumor cells
using our newly
developed
imaging device
– Hexascope.
Novel methylation-dependant
markers involved in the
progression of cancers
Recently we have developed and
used NotI microarrays (NMA).
NMA are the only existing
microarrays giving the opportunity to detect simultaneously
Eugene Zabarovsky and differentially copy number
[email protected] and methylation changes. Thus
they allow to check cancer cells
for genetic and epigenetic abnormalities. At present, we analyzed over
400 samples representing various cancers: breast, kidney, cervical,
colon, ovarian, lung, prostate, nasopharyngeal and esophageal carcinoma and leukemia. In the study 190 genes from human chromosome 3 were analyzed. For all studied cancers, we found genes specifically methylated in malignant cells. Deeper analysis of our data to find
good markers for cancer and progression detection using cervical
cancer samples revealed different methylation patterns at different
tumor stages. Increased methylation during cancer progression was,
for example, observed for the gene RBSP3 the expression of which
was subsequently shown to be downregulated. With lung cancer we
obtained similar data. Analysis of 20 prostate samples detected genes
that were not only cancer specifically methylated such as LRRC3B,
RBSP3 and GLCE, but the methylation pattern also allowed to
discriminate aggressive from non-aggressive forms of prostate cancer.
It is interesting to note
that two such progression dependant genes,
RASSF1A and RBSP3,
had an exceptionally high
incidence of single-base
mutations. Altogether in
144 sequenced RASSF1A
clones, 129 mutations
Scheme of cancer development.
were detected (mutation
frequency (MF) = 0.23 per
100 bp). In 85 sequenced RBSP3 clones, 89 mutations were found
(MF=0.10). The mutations diminished the ability of corresponding
transgenes to suppress cell and tumor growth implying a loss of function. This was the first report of high frequencies of somatic mutations
in RASSF1 and RBSP3 in different cancers. Somatic hypermutations
in tumor suppressor genes involved in major human malignancies
offer a novel insight in cancer development, progression and spread.
Selected publications
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Selected publications
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Flaberg E, Sabelstrom P, Strandh C and Szekely L. Extended Field
Laser Confocal Microscopy (EFLCM): combining automated
Gigapixel image capture with in silico virtual microscopy. BMC
Med Imaging 8, 13, 2008.
Markasz L, Skribek H, Uhlin M, Otvos R, Flaberg E, Eksborg S,
Olah E, Stuber G and Szekely L. Effect of frequently used chemotherapeutic drugs on cytotoxic activity of human cytotoxic
T-lymphocytes. J Immunother 31, 283, 2008.
Stuber G, Flaberg E, Petranyi G, Ötvös R, Rokaeus N, Kashuba E,
Wiman KG, Klein G and Szekely L. PRIMA-MET induces nucleolar translocation of Epstein-Barr virus-encoded EBNA-5 protein.
Mol Cancer 8, 23, 2009.
Department of Microbiology, Tumor and Cell Biology – MTC
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Wang F, Grigorieva EV, Li J, Senchenko VN, Pavlova TV, Anedchenko EA, Kudryavtseva AV, Tsimanis A, Angeloni D, Lerman MI,
Kashuba VI, Klein G and Zabarovsky ER. HYAL1 and HYAL2 inhibit
tumor growth in vivo but not in vitro. PLoS One 3, e3031, 2008.
Cheung AK, Lung HL, Ko JM, Cheng Y, Stanbridge EJ, Zabarovsky
ER, Nicholls JM, Chua D, Tsao SW, Guan XY and Lung ML. Chromosome 14 transfer and functional studies identify a candidate
tumor suppressor gene, mirror image polydactyly 1, in nasopharyngeal carcinoma. Proc Natl Acad Sci USA 106, 14478, 2009.
Kashuba VI, Pavlova TV, Grigorieva EV, Kutsenko A, Yenamandra SP, Li J, Wang F, Protopopov AI, Zabarovska VI, Senchenko V,
Haraldson K, Eshchenko T, Kobliakova J, Vorontsova O, Kuzmin
I, Braga E, Blinov VM, Kisselev LL, Zeng YX, Ernberg I, Lerman
MI, Klein G and Zabarovsky ER. High mutability of the tumor
suppressor genes RASSF1 and RBSP3 (CTDSPL) in cancer. PLoS
One 4, e5231, 2009.
Representatives of the technical and administrative staff at MTC. From left to right, back row: Sándor Feldötö, Greger Blomqvist, Thomas Böhlmark, John
Sennett, Åsa Sjöman, Maret Johannesson, Agneta Sandlund. Front row: Hanna Hadzija, Lada Larsson, Margareta Hagelin, Barbro Ehlin-Henriksson, Anita
Wallentin, Berit Ohlsson, Anna-Karin Persson, Milina Jovanovitch, Poonam Seth and Anna Lögdberg.
Kai Eng, Gunilla Karlsson-Hedestam and Gerrry McInerney discuss their research.
Photo: Lasse Skog
PhD student Clemens Spinnler at his annual student seminar.
Department of Microbiology, Tumor and Cell Biology – MTC
67
Avian influenza virus by Lennart Nilsson/Scanpix.
What Happens Down in the Cellar?
Lennart Nilsson at MTC
Although the Karolinska Institutet (KI) is
undermined by a network of supply corridors
there is normally not much activity going
on down there in the cellars. However, there
are some exceptions. As this environment at
MTC provides interference-free conditions
it is the ideal working place for the worldfamous photographer Lennart Nilsson and
his EOL high-resolution scanning electron
microscope. Therefore, luckily for researchers
at MTC, Lennart Nilsson has been working in
the Department for more than 5 years. Almost
naturally, this has lead to extended collaborations with some of the researchers “above”.
However, spatial vicinity has never been
a prerequisite for successful interactions.
Lennart has had a long-standing tradition
to collaborate with MTC researchers already
before the Department was founded in 1993.
He started his long-term collaboration with
Hans Wigzell, at that time professor of immu-
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Department of Microbiology, Tumor and Cell Biology – MTC
nology, in the early ’70s. Natural killer (NK)
cells of the immune system had just been
discovered and characterized by researchers at the Department of Tumor Biology led
by Georg and Eva Klein. Lennart was highly
interested in the fight for life and death
that is going on when killer cells attack and
destroy tumor cells. They also looked at blood
constituent parts, visualized viruses in the air
and discovered fossilized bacteria. Lennart
also captured infections of lymphocytes
with tumor viruses together with Ingemar
Ernberg, professor of tumor biology.
Lennart has another long-standing interaction with Britta Wahren, professor in Clinical Virology. Lennart was fascinated by the
beautiful vaccinia virus, especially how it was
released from the host cells. Some results of
the successful collaboration between Lennart
and Britta can be viewed in an exhibition at
the Swedish Institute for Infectious Disease
Lennart Nilsson. Photo: Camilla Svensk
Tumor cells attacked by spherical killer cells. By Lennart Nilsson/Scanpix.
control (SMI), among them a high-resolution
visualization of the bursting deadly virus
produced by the dying host cell.
Lennart Nilsson was among the first to
visualize the three-dimensional structure of
the highly pathogenic avian influenza virus
H5N1 at high resolution with his electron
microscope for the public. This achievement
was possible through the close co-operation
with Mia Brytting, chief microbiologist at
SMI. Their collaboration started already in
2003 with a common interest in the Severe
Acute Respiratory Syndrome (SARS) virus
shortly after the first pandemic with this virus.
To reach the general public
How does a world-famous photographer,
who already had made a world-wide career as
a photojournalist and is a sought after celebrity and court photographer, get interested in
the scientific world? Although Lennart Nilsson is grateful and very happy to work at KI,
the collaboration with researchers is actually not his major driving force. To reach the
general public, to make them see what has
never been seen before, is actually what he is
aiming for. This desire is perhaps most beautifully reflected in his famous book “A child is
born” which was first published in 1965.
To achieve his goals, Lennart has always
used front-line visualization techniques.
Starting with macro photography, he used
the scanning electron microscope. Since the
’70s. Although his own microscope can reach
a resolution of 700,000 x, Lennart is aiming
for improvements. Hitachi has built a microscope with an even higher resolution. One is
The genome seen inside the virus particle will give rise to millions of
new particles, killing the infected cell. By Lennart Nilsson/Scanpix.
located in Norway and Lennart hopes to be
able to use this equipment in the future.
Although “A picture says more than thousand words”, Lennart is also a highly appreciated lecturer who readily talks about his
work and demonstrates the techniques he is
using to anyone who is interested; may it be
researchers or school classes.
Perhaps Lennart reaches the widest audience with his films. His latest TV-production
“A journey to the core of life”, which was
produced by Mikael Agaton, was shown for
the first time in Swedish television in February 2010. It has two parts “The Cell City” and
“The Hunt for The Flu Virus”. Inspired by
Ingemar Ernbergs book ‘The ruthless cell”
(original title: Den hänsynslösa cellen), the
film was mainly produced at MTC and SMI
during 2008-2009 with the active participation of many scientists, among others Mia
Brytting and Laszlo Szekely.
Despite being 87 years old, Lennart Nilsson is involved in many new projects. Together
with Britta Wahren and Anna-Lena Hammarin, he wants to look at the rabies virus, which
has an interesting, unusual morphology.
The dangers of smoking
And most importantly, Lennart has ideas for
another educational project. A new film is
planned together with Georg Klein and his
co-workers to describe the dangers of smoking leading to lung cancer, the most common
cause of cancer-related death. Lennart wants
to visualize the transformation of a normal
lung cell into a cancer cell when exposed to
tobacco smoke for many years.
Lennart Nilsson’s fate at KI is directed
by vibrations. He and his microscope have
moved around the KI several times to find a
vibration-free location. Wherever Lennart is,
however, he continues his exciting collaboration with MTC researchers!
Ute Römling
Lennart Nilsson at KI
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In the 1970-ties Lennart Nilsson
starts to collaborate with researchers at KI.
1976 Honorary Doctorate at KI
1998 The Lennart Nilsson Award,
sponsored by the Lennart Nilsson
Foundation, honoring extraordinary photography of science was
awarded the first time.
2003 The Lennart Nilsson lecture
hall was inaugurated
2009 Personal Professorship by
the Swedish Government
2010 “A journey to the core of life”
is broadcasted on the Swedish
Tele­vision
2010 The two stamps issued by the
Swedish Post Office to celebrate
the 200th anniversary of KI depict
silicon and selenium, two of the
elements discovered by Jöns Jakob
Berzelius, one of the co-founders
of KI. The material was taken from
Berzelius own test tubes and photographed by Lennart Nilsson.
Department of Microbiology, Tumor and Cell Biology – MTC
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Foreign Adjunct Professors
Sir David Lane
p53 Forever – Focus on this
Fascinating Protein
I did my BSc in microbiology and
my PhD
in immunology both at University College
London. My first post-doctoral positions
were in virology and for most of my career
I have worked on the biology of cancer. So
MTC seems an ideal place for me to work and
collaborate with as it combines these disciplines. Increasingly I feel that breakthroughs
in research always come from crossing boundaries. It is amazing to me how easily we get
trapped into particular systems or approaches
and collaborating with others, who know
different techniques and have hands-on experience is vitally important. Immunologists
love flow cytometry and complex transplantation experiments. Molecular cell biologists
transfect everything and never mix cells of
different types together “Western blots and
HeLa cells for us!” and microbiologists? Well,
they are perfect of course and can do everything as long as it will grow in L Broth! So in
MTC I see the great power of mixing disciplines. Growing human tumor cells in Zebra
fish, examining the behavior of mice with
altered gut flora and looking at the anti-parasite activity of new anti-cancer drugs are all
being tried here.
If the new system is two buildings away
and established by someone you don’t know
the casual but vital “let’s just try it” conversation over coffee with a colleague is less likely to
happen. Of course, what everyone wants these
days is to plan science and calculate outcomes
in advance. How boring if we know what we
are going to discover in advance! How will we
really discover anything new? But of course
once the initial observation is made then the
work begins; the work to make the results
certain, quantitative and reproducible by the
community so that they have real value. Then
we can write the paper. Its odd talking to some
scientists now that the paper has become a
painfully produced product. It should never
be that way. When we have a great result the
paper writes itself.
My first contacts with MTC were, of course,
through Georg and Eva Klein’s lab where Georg
developed a great interest in the interactions
of the EBV virus proteins with p53, the most
frequently mutated gene in human cancer. At
that time (the early 1980’s) it was becoming
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Department of Microbiology, Tumor and Cell Biology – MTC
clear that all of the DNA viruses might inactivate p53 function if successful in infection of
transforming mammalian cells as the replication of viral DNA acts as a trigger to induce the
p53 response and this potentially blocks infection and transformation. While great progress
was made on this problem with discovery of
p53 binding and inactivating proteins in the
SV40 Adenovirus and Papilloma virus systems
the results where much less clear in the case of
other viral groups notably EBV, other Herpes
viruses and Polyoma virus. Like so many others
Georg “adopted” me and I was asked to act as
opponent on the thesis exams of some now
very notable MTCers. So many years later it is
wonderful to be able to join the Department on
a more formal basis. It is also very fitting that
Georg and his team now seem to have solved
the route by which EBV tackles p53.
Since the role of p53 mutations in human
cancer became apparent in the early ’90s work
on this fascinating protein and the mechanisms by which it is controlled have been
the main focus of my work though I love to
play with other things too. We still have work
going on RNA helicases, the proliferating cell
nuclear antigen PCNA, eukaryotic translation
initiation factor 4E and antibodies though not
on SV40T antigen anymore. I love proteins
and antibodies and trying to make things, so
drug discovery attempts have been a big part
of the last decade. I set up a biotech company
(Cyclacel) that progressed three molecules to
the clinic and I was heavily involved in science
policy and development with A*Star, the
agency for science, technology and research in
Singapore and Cancer Research UK CRUK, a
main funder of cancer research in Britain.
But now I am back to looking at bands
on gels, making and testing antibodies and
screening libraries looking out for amazing
surprises. So what do I hope for the interaction with MTC? Lots of great discussions over
coffee, lots of new ideas and collaborations
and lots of fun. I had great mentors when I
was a student, especially my PhD supervisor
Avrion Mitchison, who was very inspiring
and extra ordinarily generous. So I would like
to be like him and help all the students when
I can.
Current positions
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Director p53 lab Singapore 2008–
Chief Scientist A*Star Singapore 2008–
Chief Scientist, Cancer Research UK
2008–
Foreign adjunct professor at the Karolinska Institutet/MTC 2007–
Honorary Professor, University of
Dundee, 2009–
Personal information
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Research interests: Original discovery of p53
and subsequent research on p53 all the way
from a basic research to clinical application
for three decades, over 340 publications
among others in Nature, Cell and Lancet.
Prizes (selected): Paul Ehrlich Prize, 1998;
Knighthood, 2000; Sergio Lombroso Award
in Cancer Research, Weizmann Institute
of Science, 2005; Hon. Doctor of Science,
University Paul Sabatier, Toulouse; University of Nottingham; University of Birmingham; University of Aberdeen and others.
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PhD in immunology, University College
London, Great Britain
1977–1985 Lecturer, Imperial College of
Science & Technology, London
1985–1990 Senior and Principal Staff
Scientist, Imperial Cancer Research Fund,
Clare Hall Laboratories
1990–1998 Department of Biochemistry,
University of Dundee
1990–2009 Personal Chair in Molecular
Oncology, University of Dundee.
1998–2009 Department of Surgery
and Molecular Oncology, University of
Dundee
1998–2009 Director of the Cancer
Research UK Cell Transformation
Research Group, The Cancer Research UK
Laboratories at Dundee
2004–2007 Executive Director of the
IMCB in Singapore on a three year leave
2006–2008 Founder of the Experimental
Therapeutics Center (ETC) Singapore
2008–2009 Chair of Singapore’s Biomedical Research Council
Foreign Adjunct Professors
Margret Ann Liu
MTC – Not Only a Scientific
Experience!
My work focuses on global health, particu-
larly, on vaccines and immunotherapy for
diseases such as AIDS and cancer. Being
initially a Visiting Professor and now a Foreign
Adjunct Professor at the Karolinska Institutet (KI) in MTC has provided a wonderful
involvement in ongoing projects and engaging interactions with the faculty and students.
My activities have ranged from teaching
in courses, to co-supervising PhD students
to being a thesis opponent, to participating in
the various European and developing country networks to which MTC and the nextdoor Swedish Institute for Infectious Disease
Control (SMI) belong. Applying for research
grants is another activity which I had not done
since my early days on the faculty at Harvard
Medical School, and which has been quite a
change from my days as the Senior Advisor
in Vaccinology for the Bill & Melinda Gates
Foundation overseeing many millions of
dollars of funding for vaccine development!
Vaccine development
My primary affiliation is with the research
group of Britta Wahren, currently Professor
Emerita, and formerly Professor of Virology
at MTC/SMI, developing a vaccine for HIV/
AIDS and immunotherapy for cancer, collaborating also with Prof. Håkan Mellstedt, Chief
Physician, Departments of Oncology and
Hematology, Karolinska University Hospital.
These projects are particularly rewarding for
me because through our efforts to improve
DNA vaccines (an approach my former
research group and I pioneered in the 1990s
at Merck Research Labs), our KI group and KI
clinicians are trying to benefit humankind by
testing an HIV vaccine and a cancer immunotherapy in clinical trials.
In fact, my appointment at the KI
stemmed from this research as I was first
introduced to Professor Wahren’s research
and group at meetings, and then in 2001
gave an honorary lecture in the Karolinska
Research Lecture series at the invitation of
the Nobel Committee. The internationally
collaborative research of the KI scientists has
complemented and expanded the work that
I do for a variety of international organizations focused on improving global health by
developing vaccines needed by the world’s
poorest people. For example, for the International Vaccine Institute (IVI), established by
the UN Development Program, and located
in South Korea, I was the long-time Chair of
the Scientific Advisory Group, and now serve
as Vice-Chairman of the Board of Trustees,
coincidently under the Chairmanship of
Prof. Ragnar Norrby, who recently retired
from his position as Director General of SMI.
The IVI recently enabled the production
and licensure of a low-cost cholera vaccine
to prevent untold suffering and mortality in
developing countries. Likewise, when I have
organized international meetings, whether
in the US, Asia or Africa, KI faculty, students,
and post-docs have been speakers and participants. One encounters Karolinska scientists
throughout the world, so it is optimal to also
work with them in Stockholm!
Personal information
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Research interests: DNA based vaccine
development, immunotherapy; inventor of six patents; organizer of a dozen of
international meetings among them Cold
Spring Harbor Laboratory meetings and
Keystone Symposia.
Co-Editor of the 3rd and 4th editions of
“New Generation Vaccines”.
Prizes: Recipient of an NIH Physician
Scientist Award; Named 1 of “The 50 Most
Important Women in Science” by Discover
magazine, 2002.
Memberships (selected): Vice-Chairman of
the Board of Trustees, International Vaccine
Institute; Director, Keystone Symposia
Board; Trustee, San Francisco Conservatory of Music; Advisory Board, Elizabeth
Glazer Pediatric AIDS Foundation.
A new social life
Being part of MTC and the KI has also
resulted in friendships, cross-cultural sharing, and a huge improvement in my social life!
I have been privileged to attend the incredible
Nobel Prize ceremony and dinner as an affiliated faculty member of the KI. Also no parties
elsewhere can compare with MTC parties.
Who can forget Prof. Ingemar Ernberg playing the White Rabbit in an “Alice in Wonderland” satirical skit, complete with ears and
hopping around the stage, demonstrating
that a department chairman has many hidden
talents.
Another outcome of these parties and
thesis celebrations is that my extremely
limited Swedish vocabulary was expanded by
singing drinking songs. Professor Wahren has
welcomed even my family to stay in her home,
providing us with an opportunity to discover
the wonders of Sweden. What can I say, but
“Tack så mycket!” for the incredible experience of being part of MTC and the KI.
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Current positions:
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ProTherImmune, Consultant in Vaccines
and Immunotherapy for Biotech and
Investment Companies, 2000–
Foreign Adjunct Professor, KI, Stockholm,
Sweden, 2007–
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1977 B. A. in Chemistry, Colorado College;
1977 Teaching diploma for piano, Ecole
Normale de Musique de Paris
1981 M.D., Harvard Medical School,
Boston, MA
1981–1988 Internship, Residency in Internal Medicine and Clinical and Research
fellow, Massachusetts General Hospital,
Boston, MA
1984–1988 Visiting Scientist at the Massachusetts Institute of Technology, Boston,
MA
1987–1989 Instructor in Medicine at
Harvard Medical School, Boston, MA
1988–1997 Senior research fellow, later
Director and Senior Director, Merck
Research Laboratories, West Point, PA
1989–1995, Adjunct Assistant Professor
of Medicine, University of Pennsylvania,
Philadelphia, PA
1997–2000 Vice President, Vaccines
Research, Chiron Corporation, Emeryville, CA
2000–2006 Vice Chairman, Transgène,
Strasbourg, France
2000–2003 Senior Advisor in Vaccinology and Consultant, Bill & Melinda Gates
Foundation
Department of Microbiology, Tumor and Cell Biology – MTC
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Foreign Adjunct Professors
Alexander von Gabain
Innovating Vaccines by
Translational Entrepreneurship
Vaccination is inarguably the most success-
ful medical intervention which has become
a mandatory part of many countries’ health
care programs during the last century and
has shown to be an effective instrument in
the control of infectious diseases worldwide.
Development and launch of novel vaccines
has been triggered by the appearance of
novel pathogens, by the apparent stagnation
to control the rebound of global infectious
diseases, by the pending threat of a pandemic
flu and by the spread of multi-drug resistant
pathogens in hospitals and the community.
The encouraging scientific progress made in
the areas of immunology, molecular biology,
genomics, and host-parasite interaction, but
also in the areas of novel manufacturing technologies, has facilitated the development of
novel vaccines and enabled the field to expand
the spectrum of the existing vaccine targets.
Innovation in vaccine development is
mostly driven by small and medium size
biotech enterprises. The classical path giving
rise to novel biotech firms has been the spinoff from an academic environment; e. g.
university scientists stepping out of their
institutes, converting into entrepreneurs
and homing their scientific knowledge and
research results into a business shell, aiming
to translate the founding concept and vision
into viable and badly needed products. This
approach has led in single instances to the
foundation of impressive companies, some
of which have outgrown established pharmaceutical companies.
The Company’s growth is nurtured by
three pivotal platforms forming the fundament of Intercell: the Antigen Identification
Program® enabling the discovery of the most
protective microbial antigens, the vaccine
adjuvant programs that increase the efficacy
of existing and novel vaccines, and the patch
delivery technology that pioneers a needlefree era in the vaccine field. Based on these
platforms, the Company was able to build a
coherent Research & Development pipeline
that today comprises more than eight products in various stages, currently tested in
Phase I to Phase III clinical trials. The Company’s most advanced product, a novel prophylactic vaccine against Japanese Encephalitis,
was launched after its global registration
in 2009. For more information please visit:
www.intercell.com.
Since my appointment as foreign adjunct
professor, I have been engaged in numerous activities aiming to transmit and share
my knowledge, networks and experience in
microbiology, vaccinology, but also in biotech
entrepreneurship and translational medicine
with the MTC students, MTC colleagues and
other Karolinska staff. Among other activities,
I have supported the setup of collaborations
between MTC and Intercell scientists on novel
vaccines over the last decade, but also organized a teaching course on infectious disease
biology including PhD students from Vienna
and Stockholm. Furthermore, I am coaching
students and faculty to spin off companies
from the Karolinska Institutet (KI) with its
impressive foundation of top class research.
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von Gabain A. Innovating vaccines: from
gene expression to translational entrepreneurship. Hum Vaccin 6, 1, 2010.
Current positions
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Chair of the Scientific Advisory Board and
Strategic Advisor to the Board Intercell AG,
2009–
Professor in Microbiology at the Max
Perutz Laboratories, University of Vienna,
Austria
Foreign Adjunct Professor at the KI/MTC,
2007–
Personal information
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Research interests: Microbial gene expression, host-parasite interactions and immunology. Over 100 articles in peer-reviewed
journals including Nature and Cell. Regularly speaks at and organizes conferences in
the field of vaccinology.
Prizes: Scrip Award with Intercell AB as
Biotech Company of the Year 2008
Memberships (selected): Member of the
Royal Swedish Academy of Engineering
Sciences, the founding governing Board
of the European Institute of Technology
(EIT); board member of biotech enterprises including TVM Capital in Munich,
Germany
Foundation of Intercell
Inspired by this paradigm and by the ambition to translate research into novel desperately needed vaccines, I stepped down from
my academic Chair at the Campus Vienna
Biocenter (VBC, for more information please
visit: http://www.viennabiocenter.org) and,
together with four colleagues, founded Intercell in 1998. As founder and in various board
positions, I have been contributing to leading
the Company into the top league of the 30
largest biotech firms worldwide, with today
450 employees in Austria, the UK and the
USA, and technology provider to the five largest commercial vaccine players.
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Department of Microbiology, Tumor and Cell Biology – MTC
Selected publications
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Giefing C et al., Discovery of a novel class
of highly conserved scale antigenic fingerprinting of pneumococcus with human
antibodies. J Exp Med 205, 117, 2008.
Kamath AT et al., Protective anti-mycobacterial T cell responses through exquisite in vivo activation of vaccine-targeted
dendritic cells. Eur J Immunol 38, 1247,
2008.
Schlick P et al., Helices a2 and a3 of West
Nile virus capsid protein are dispensable
for assembly of infectious virions. J Virol
83, 5581, 2009.
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PhD in Genetics, University of Heidelberg
1979–1982 post-doc in the laboratory of
Stanley N Cohen, University of Stanford
1983–1992 Associate Professor, University
of Umeå and KI
1993–1998 Head of Department of the
Microbiology and Genetics, University of
Vienna, Campus Vienna Biocenter
1998–2005 Co-founder and CEO of the
biotech company Intercell AG
2005–2009 CSO and chair of SAB, Intercell AG
Foreign Adjunct Professors
Yi-Xin Zeng
Our valuable Collaboration
with the Karolinska Institutet
the immunotherapy of nasopharyngeal carcinoma, and together we have published several
East Indian Company tradeship “Götheborg”
important papers in the field.
that had returned to China 265 years after its
Another collaborative effort with great
latest journey, I saluted Her Royal Highness
impact is the China-Sweden Medical SympoQueen Silvia in Guangzhou. Meanwhile, the
sium. In 2003, the 1st China-Sweden Medical
3rd China-Sweden Medical Symposium was
Symposium was held in SYSU cancer center.
held in the Sun Yat-Sen University Cancer
In 2004, the 2nd China-Sweden Conference
Center. At the opening ceremony, I had the
was held in Stockholm. Accidentally, the
honor to receive the official certificate for my
Nobel Prizes were awarded during the time the
appointment as a foreign adjunct professor at
conference took place. As
MTC, Karolinska Institutet
a Chinese representative,
(KI), from Professor Jan
I was invited to particiCarlstedt-Duke, the Dean
“I really cherish the great
pate in the ceremony for
of Research at that time. It
honor as a foreign adjunct
the Nobel Prizes, which
was an exciting moment,
professor of KI and will
left a life-long memory
and many distinguished
work as hard as always to
to me. In 2006, the 3rd
guests witnessed and shared
achieve more in scientific
China-Sweden Medical
my happiness, including
research and medical
Symposium was again
Her Royal Highness Queen
education and management
held in the SYSU cancer
Silvia, the Minister of
as well.”
center. During these
Science and Technology of
conferences,
experts
China, Professor Guanhua
from both China and Sweden reported their
Xu, and the Vice President of Chinese Acadnovel achievements and intriguing discusemy of Sciences, Professor Zhu Chen who is
sions about different aspects of biomedicine
now the Minister of Health in China.
were initiated, which eventually promoted
collaborations among scientists between the
Many collaborative efforts
two countries.
Since 2003, when the 1st China-Sweden
I really cherish the great honor as a foreign
Medical Symposium was held in Guangzhou,
adjunct professor of KI and will work as hard
Sun Yat-Sen University (SYSU) has cooperas always to achieve more in scientific research
ated with KI in a range of aspects in biomediand medical education and management as
cal research. On March 27, 2004, the SYSU-KI
well. This year will be the 200 anniversary of
collaborative laboratory was established. The
KI and I hope I can join the celebration and
collaboration with KI has three goals: trainexpress my best wishes for KI.
ing of PhD students or post-doctoral research
who will most likely return to China; the
application of joint grants and the formation
Current position
n Director of the State Key Laboratory of
of joint projects of mutual interest with Swedish partners. As a foreign adjunct professor at
Oncology in Southern China, 2004–.
n Professor and President of the Sun YatKI, I have obtained a large number of benefits that arise from it, such as training PhD
sen University Cancer Center, the largstudents. For example, Roujun Peng, one of
est specialized Cancer center in Southern
my PhD students, is doing her PhD research
China, where cancer treatment, cancer
on the subject of immunological genetics at
prevention, research and training are inteKI right now.
grated, 1997–.
n Foreign Adjunct Professor, KI, Sweden,
Another benefit is the cooperation in the
area of cancer immunotherapy, which has
2007–.
produced a great deal of achievements in the
past three years. Jiang Li, a research scientist
Personal information
n Research interest: Cancer immunotherain my group, collaborated with Prof. Maria
Masucci, professor in virology, focusing on
py, Cancer genetics, nasopharyngeal carIn the summer of 2006, with the arrival of the
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cinoma and Epstein-Barr virus associated
malignancies.
Prizes: Award for Outstanding Young
Teachers in Chinese Universities, Ministry
of Education, 2002; First class award for
Scientific Advancement, China Medical
Association, 2003; Second class award for
Natural Science Achievements, National
Office for Science and Technology Awards,
2005; Award for National Young Scientists,
2006; He-Liang-He-Li Award for Science
and Technology Advancement, 2007.
Memberships (selected): Member of the
Chinese Academy of Sciences; Member
of the Academy of Sciences of Developing Countries, Member of the SFDA
(Sino-Food & Drug Administration)
Drug Evaluation Committee. Editor-inchief, Chinese Journal of Cancer; Editorial Board member of Cell Cycle (USA),
Cancer Biology and Therapy (USA), Journal of Biological Chemistry (USA) and
Journal of Translational Medicine (USA).
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1990 PhD at the Sun Yat-sen University of
Medical Science, China
1990–1992 Research Assistant, GuangDong Provincial Peoples Hospital, China
1992–1994 Postdoctoral Fellow at the
Institute of Medical Science, Tokyo
University and Tokyo Metropolitan Institute of Gerontology, Japan
1994–1997 Research Associate, Howard
Hughes Medical Institute, University of
Pennsylvania School of Medicine, USA
1997 Professor and Vice President, SunYatsen University Cancer Center, China
Department of Microbiology, Tumor and Cell Biology – MTC
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MTC International
Evaluation of HIV Vaccines in Tanzania
Development of promising vaccination strategies at MTC and SMI
Karolinska Institutet (KI) and the Swedish
Institute for Infectious Disease Control (SMI)
have for many years had a Sida-supported
research collaboration with the Muhimbili University of Health and Allied Sciences
(MUHAS) in Dar es Salaam, Tanzania called
the TANSWED program. This program
comprises studies of various aspects of HIV
infection in Tanzania. An important part of
this program is the building of research capacity which includes PhD research training at
KI of medical doctors from MUHAS. One of
the main projects in the TANSWED program
concerns “Evaluation of HIV vaccine candidates”.
After more than 10 years of preparatory work a HIV vaccine trial (HIVIS03)
supported by Sida and EU started in Dar es
Salaam in 2007 using a combination of two
HIV vaccines. The first vaccine is an HIV-1
subtypes A, B and C. In order
to augment and broaden the
vaccine-induced
immune
responses a prime-boost
vaccine strategy has been
implemented. This means that
boosting immunizations were
given with a modified vaccinia
virus Ankara (MVA) vectorbased HIV vaccine representing subtypes A and E. The HIV
MVA vaccine was obtained
through collaboration with the
National Institutes of Health
HIV DNA vaccination of the first volunteer, a police officer, in the
and Walter Reed Army InstiHIVIS03 vaccine trial in Dar es Salaam, using a needle-free injection system.
tute of Research in the US.
This HIV vaccine combination was first tested in a clinical trial of 37
DNA vaccine developed at KI/SMI which
healthy non-infected vaccine recipients in
consists of genes representing the major HIV
Stockholm (HIVIS 01/02) coordinated by Eric
structural proteins and three major HIV
Participants in the annual HIVIS/TaMoVac HIV vaccine meeting in Bagamoyo in Tanzania including collaborators from Tanzania, Mozambique, Sweden, Germany, England and the US.
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Department of Microbiology, Tumor and Cell Biology – MTC
MTC International
Sandström. The vaccine combination was
shown to be safe and highly immunogenic. All
vaccine recipients except one showed an HIVspecific cellular immune response.
The positive HIV vaccine results in Stockholm have been confirmed in the HIVIS03
vaccine trial in Dar es Salaam which included
60 volunteers from a cohort of police officers. After 3 HIV DNA vaccinations followed
by one HIV MVA vaccination all of the 35
vaccine recipients displayed HIV-specific
cellular immune responses. It was found that
a lower dose of HIV DNA vaccine given intradermally induced stronger immune responses
after the HIV MVA boosting vaccination than
did a higher HIV DNA dose injected intramuscularly. One reason for the excellent
immunogenicity is that the delivery of the
vaccine genes occurs via a jet stream using a
needle-free injection system. The use of this
technology involved collaboration with the
Bioject company in the US. Placebo recipients
showed no positive responses.
Participants on the ”Genes and Genomes” in the Tropics course.
Scientific Alliances in Africa
are involved in
numerous research and teaching efforts
throughout Africa; courses are given and
scientific alliances have been formed.
We are fully connected and collaborate
both for the sake of Swedish and African
science. Makerere University in Uganda,
(MUK) in particular, has an outstanding relationship with KI for many years.
The signing of a contract of collaboration
in 2002 by the heads of the two institutions formally established the importance of the inter-university links. We
have as a consequence a joint degree for
PhD students, unique in the world, with
43 graduates so far. Critical findings have
been reported in outstanding scientific
journals and our collaborative activities
have changed both KI and MUK.
At Makerere University a collaborative
three-weeks PhD course named “Genes
and Genomes in the Tropics” (www.
ggtropics.com) is executed on an annual
basis. It was created in the year 2000 in
an attempt to increase the knowledge of
front-line molecular- and cell biology of
microbes but also to stimulate collaboration and networking between the two
universities. 175 students of 15 nationalities, predominantly from KI and MUK
have taken the course, but students and
teachers have also come from Cameroon,
Denmark, Ethiopia, France, Germany,
India, Italy, Kenya, Mali, Nigeria, Norway,
Portugal, Sudan, Tanzania and USA.
Scientists at MTC
Two new vaccine projects
Funding has been obtained from the European and Developing Countries Clinical
Trials Partnership (EDCTP) and Sida for
two new big vaccine projects, TaMoVac I and
TaMoVac II. These projects include building
of HIV vaccine trial capacity and conduct of
phase II HIV vaccine trials in Tanzania and
Mozambique with the use of the HIVIS HIV
DNA and MVA vaccines.
These trials will evaluate ways of facilitating the injection of the HIV DNA vaccine
and of enhancing the induction of cellular immune responses as well as antibody
responses to HIV. A new concept will be
introduced with the delivery of the HIV DNA
genes by dermal electroporation. In this way
we hope to further increase the vaccine gene
influx into dermal cells and the expression of
viral proteins. Preclinical studies have shown
that this will considerably increase immunogenicity.
The principal investigators of the TaMoVac I and II vaccine projects are two Tanzanian doctors who took their PhD degrees at
KI, Muhammad Bakari, a clinician and Eligius
Lyamuya, a clinical immunologist. Five Tanzanian and two Mozambican PhD students are
presently engaged in these projects. Three
of them are already registered at KI for PhD
studies. The collaboration includes investigators from Tanzania, Mozambique, Sweden,
Germany, England and the US.
Gunnel Biberfeld
Our research is focused on diseases
such as malaria, Burkitt’s lymphoma,
AIDS, tuberculosis and urinary tract
infections. More than 20 separate studies have been carried out and published
in collaboration between KI-MTC and
MUK. For example, the ability of the
malaria parasite Plasmodium falciparum
to undergo antigenic variation has been
investigated and a scheme was identified in the variable antigens pertinent to
disease states and parasite phenotypes.
In this and the other projects, PhD
students and scientists from Stockholm
and Kampala have worked at KI-MTC
and MUK. Fellow students, “mirror
students”, from Uganda or from Sweden
devoted to the work with the same overall question and the same timeline have
been engaged in the projects in order to
install reciprocity in the support and in
the work. This has allowed for parallel
help and development from Uganda to
Stockholm and vice versa.
The close interaction seeks to avail
opportunities for productive research
and training of both young Ugandan
and Swedish students. This has had both
scientific and societal impact fulfilling
social and economic needs by creating
knowledge and empowerment. We live
in a world developing at a rapid pace
but research still needs to be a long-term
activity.
Mats Wahlgren
Britta Wahren
Department of Microbiology, Tumor and Cell Biology – MTC
75
MTC International
Singapore skyline. Photo: bigfoto.com
The Singapore Partnership for the Future
The country of Singapore, located in the
South East of Asia, with 4.5 million citizens is
a vibrant modern cosmopolitan country with
a strong economy and well developed infrastructure to support Research and Development (R&D). In addition, Singapore is a
harmonic introduction and gateway to eastern culture and values. Since the end of 2002,
I have been given the privilege to coordinate
scientific research and education exchange
activities between Karolinska Institutet (KI)
and Singapore. In the early 2010, KI has well
developed programs and tight bonds with
the Universities of Singapore (NUS) and
Nanyang Technology University (NTU) as
well as the Minister of Trade funded Singapore Agency for Science, Technology and
Research (A*STAR).
Over the last five years Singapore has
established itself as a major player in R&D
both in South East Asia as well as internationally. Singapore can not compete in size
and numbers with other major players such
as India and China. Yet, its considerable
investment in Life Science Research and its
geographical location in the heart of South
East Asia, make Singapore a very attractive
partner for KI and Sweden.
76
Department of Microbiology, Tumor and Cell Biology – MTC
Singapore and Sweden
Singapore is an internationally competitive
country eager to foster international collaborations with trustworthy and committed partners. Sweden is a solid partner for Singapore.
Both countries are short of human capital –
i.e. we are both in need of a constant influx
of highly educated immigrants. Joint activities are therefore of mutual benefit. Partnership with Singapore is giving KI and Sweden
access to intellectual capital, which already is
becoming a strong competitive advantage at
the global R&D arena.
Furthermore, as English is the main
spoken language, no communication barriers
exist. Given the existence of good instruments
to handle ethnic pluralism, Singapore has a
lot to offer to KI and Sweden in terms of adaptation and learning about other ethnic values,
information that is useful for our efforts to
work and operate on the international arena,
be it Europe, US or Asia.
Singapore and KI
Currently KI is engaged in PhD programs
with the three major actors, A*STAR, NUS
and NTU respectively. The most productive
program so far is the one with A*STAR. For
more information see: http://newshub.nus.
edu.sg/ke and the KI homepage.
Other examples of exchange programs
are the possibility for PhD students from KI
to visit A*STAR research institutes at Biopolis for up to one year. These programs are
sponsored by the generous support from the
universities and A*STAR.
An overseas office for KI was opened in
Singapore in 2004. This office has since then
been used to establish a trust and respect for
our mutual efforts to work and operate in
Singapore.
Large scale collaborative research activities are ongoing in the areas of breast cancer
and Chronic Inflammatory Diseases (CID),
health care problems of huge magnitude
both in Sweden and in Singapore. Scanning
the entire human genome for genes predisposing to CIDs is very costly, technologically demanding and requires a well defined
clinical material. In recent years, the A*STAR
funded Genome Institute of Singapore (GIS)
and KI have successfully collaborated and
identified biomarkers for multiple CIDs
including rheumatoid arthritis, psoriasis
and ulcerative colitis. It is expected that additional research exchange project in other
MTC International
areas will be launched during the next couple
of years.
Promoting Sweden - The Sweden Day
To promote and foster interactions at the
undergraduate level, we launched the
“Sweden Day” in 2006. This event is held
twice a year and is organized by undergraduate students from KI and other Swedish
university students visiting Singapore. Their
mission is to act like “mini diplomats” and
promote higher education possibilities in
Sweden in general and KI in particular. The
students work closely with staff from the
Swedish Embassy. For further information,
see: http://swedenday.bloggplatsen.se/ where
you can read more about our activities and
the latest Sweden Day.
Singapore 2010 and beyond
Karolinska Institutet needs strong partners
to be internationally competitive if we are
to remain a major player in R&D in the field
of Life Science. R&D today does not respect
national borders, requires solid infrastructures and a long term commitment by the
politicians to be competitive. Asia has huge
unmet needs for better health care and the
investment in medical research will therefore
continue to increase in decades to come. This
makes Asia very attractive to KI and Sweden.
The build up of strong ties with Singapore is
therefore essential as well as those currently
under way with distinct regions in China and
India. KI has a good reputation in Singapore
and the KI brand is well respected for its R&D
in translational research.
International networks for
high-quality graduate education
MTC has strong international connections. The effective networking is also
reflected in the graduate education profile
of the Department. Within the field of
Infection Biology, several research groups
at MTC participate in two networks for
graduate education within Europe:
1. EIMID – European Institute of Microbiology and Infectious Diseases – is a
collaborative research initiative between
five leading European institutions in the
fields of Microbiology and Infectious
Diseases; the Max-Planck Institute for
Infectious Biology in Berlin, Germany;
the Centre for Molecular Microbiology and Infection (CMMI) at the Imperial College in London, UK; the Institute
Pasteur in Paris, France; MTC at the
Karolinska Institutet (KI) and Novartis
Vaccines and Diagnostics in Siena, Italy.
EIMID conferences have been held at
different geographic locations once a year
since the start in 2004.
2. The European Research Training Group “IRTG 1273 Strategies of
human pathogens to establish acute
and chronic infections” is a collabora-
tive network between Hannover Medical
School (MHH), the TWINCORE Institute for experimental and clinical infection research in Hannover, the Helmholtz
Centre for Infection Research in Braunschweig, and the Karolinska Institutet
in Stockholm. The central aim of this
program is to give young researchers from
Germany and Sweden the opportunity
to perform their PhD thesis in the field
of infection biology and human microbial pathogens in a highly stimulating,
competitive and international environment. The core element of IRTG 1273 is
a joint research program centering on the
question how human microbial pathogens achieve acute or persistent infection and how the host organism reacts to
infection. Twenty-four research groups
and twenty-seven graduate students
from several institutions at KI have been
involved in this program. Common
meetings have been organized within the
program at least once a year in Germany
and Sweden and students have visited
the other country to perform common
projects in collaborative efforts.
Birgitta Henriques Normark
Mikael Rhen
A long lasting friendship
KI has made a firm commitment to its senior
representatives in the Life Science program,
to develop this partnership into a long lasting friendship. Long term objectives for
both parties are to deliver first class research
coupled with excellent education programs
for students, scientists and for people working
in the health care sector. The growing interest from many other Swedish universities to
participate in our ongoing R&D activities in
Singapore together with the well developed
Swedish private industry operating in Singapore, open for more exciting investments in
Singapore in the years to come.
The upcoming bilateral national R&D
exchange program between Sweden and
Singapore, to be signed off in the near future,
make this journey very exciting.
Sven Pettersson
Department of Microbiology, Tumor and Cell Biology – MTC
77
Scientific Networks
Scientific Networks
KICancer retreat September 2008 at Utsikten, Nynäshamn.
KICancer
KICancer is a global network for all
research groups active totally or partly in
cancer research at the Karolinska Institutet
(KI), from clinical to preclinical science and
from epidemiology to nursing care. Almost all
groups working in the area have now joined
the network, therefore KICancer gathers
close to 130 groups. This accumulation of
competence makes cancer research the largest
research discipline of KI.
KICancer was initiated in 2003 as the first
translational network at KI; the need to closer
connect clinical with preclinical competence
and to establish common research projects
was especially obvious in this research area.
Now eleven networks covering the major
research areas of KI are established. The
idea of translational networks was already
launched in 1996 by the dean of research and
professor of virology, Erling Norrby, but it did
not take off at that time.
KICancer has adopted four major missions:
1. To offer “meeting places” in a broad sense,
on the Web, http://researchnetworks.ki.se/
converis//area/182, in seminars, at conferences and the annual, much esteemed KICancer retreat, usually held at Djurönäset in the
Stockholm archipelago.
2. To catalyze novel, translational research
projects and initiatives. This was achieved
by the award of grants such as the Strategic
Grants; 8.5 million Swedish crowns were
awarded to nine interdisciplinary, interdepartmental projects. In addition, eleven
diagnose-based networks were initiated since
2008 to catalyze interdisciplinary collaboration around the major cancer forms.
Douglas Hanahan, a member of the KICancer
International Advisory Board, talks at the KICancer
retreat.
78
Department of Microbiology, Tumor and Cell Biology – MTC
3. To organize the Research Training program
in Oncology and Tumor Biology, offering
around 15 courses/year for graduate students.
4. To assist in establishing networks and
collaborations with major cancer centers in
the world. Joint programs have been initiated with German Cancer Research Center
(DKFZ) in Heidelberg, the Netherlands
Cancer Institute (NKI) in Amsterdam, the
Institute Gustave-Roussy (IGR) in Paris
and the Harvard Cancer Center (including
Danna-Farber) in Boston. Collaborations
with Cancer Centers in Guangzhou and Tianjin, China have also been established.
An international Advisory Board
In 2009, the network was delighted to appoint
a prominent International Advisory Board
with David Livingston, Danna-Farber,
Boston; Doug Hanahan, Lausanne; Mina
Bissell, Lawrence-Berkeley and the Nobel
laureate of 2008 Harald zur Hausen, Heidelberg, Germany.
KICancer has catalyzed and inspired
many new collaborations in the field. It has
stimulated close interactions between cancer
researchers at KI and provides a common
voice to the KI leadership and the world
outside of KI, such as the general public,
companies and other universities.
Ingemar Ernberg, Chairman
Scientific Networks
KI Inflammation
and Immunology
The KI Infection Board Members 2006 from left to right: Mikael Rhen, Sven Britton, Gunilla Karlsson Hedestam, Mats Kalin, Staffan Normark, Kristina Broliden, Markus Maeurer, Anna-Lena
Spetz and Jan Albert.
KI-Infection
The KI-Infection Network has been up
and running since 2005 and now has over
200 members including both established
researchers and students at the Karolinska Institutet (KI). The Network aims to
strengthen infectious disease research at
the KI by promoting interactions between
scientists working in all disciplines of infectious diseases. In addition the Network acts
as a hub for infection biology-related information and highlights interesting news and
events within the subject.
A regular KI-Infection e-newsletter is
sent to all members of the Network and the
KI-Infection homepage (http://researchnetworks.ki.se/converis/area/187) is the
central reference point for the Network’s
activities. The KI-Infection Network has
members from many departments, including MTC, the Center of Infectious Medicine (CIM), Department of Medicine and
Clinical Microbiology at Karolinska Hospital, amongst others. In 2009 a new steering group was established consisting of
clinicians and non-clinical researchers in
multiple disciplines including bacteriology,
virology, parasitology and immunology.
The already existing and well functioning Postgraduate Program in Infection
Biology, dedicated to the education of
PhD students, has now become an integral
part of KI Infection. The Study Counselor
for the Postgraduate Program is Benedict
Chambers (CIM) and the Program Director is Mikael Rhen (MTC). Each year the
Postgraduate Program in Infection Biology
and the KI Infection Network runs a retreat
outside of Stockholm. In 2009, the retreat
was held in Gothenburg, well attended by
researchers within the Infection Biology
Network at KI, as well as researchers within
infection biology at Stockholm University, Uppsala University and the Hannover
Medical School, Germany. This intensive
collaboration between KI and Hannover
exists as part of common international
graduate program. In addition to the
retreat, the Postgraduate Program in Infection Biology also coordinates an annual
one-day retreat at Norra Latin in Stockholm
where PhD students present their work.
In 2010 the KI-Infection network will
recruit new members with a particular
focus on attracting clinical researchers and
strengthen ties between the various subjects
in infection research.
Mikael Rhen, Chairman
Laura Plant, Coordinator
The Karolinska Institutet (KI) network in
inflammation and immunology, KiiM, was
formed in January 2005. The idea behind KiiM
was to find a platform for interaction across
department borders, to facilitate exchange of
ideas and to inspire scientific discussions and
collaborations, but also to serve a very practical
site, namely to exchange methods and reagents.
KiiM arranges seminars where internationally
renowned scientists are invited, and where KiiM
members meet and discuss science.
KiiM also arranges a biannual highly appreciated retreat, which usually attracts 100 students,
post docs and senior researchers. The latest one,
held in October 2009, included, besides a few
invited speakers, a very successful poster session
and many impressive student presentations. In
addition, a bench-to-bedside discussion on the
anti-inflammatory effects of the lipid lowering statins was held. The retreat was a social and
scientific success; many new collaborations were
formed. KiiM is also building a project database
and manages a very active email list which is used
for search of reagents, collaborations and as a
means of information about activities of interest
for KiiMers. Many activities are arranged in collaboration with the KI Doctoral program Aii (Allergy
Immunology and Inflammation) and with CfA
(the Centre for Allergy Research). Together, these
three organizations represent most KI researchers,
from doctoral students to professors, within the
field of immunology and inflammation.
KiiM is headed by a steering committee of 15
people representing many different departments
and areas of preclinical as well as clinical research.
The steering group is chaired by Petter Höglund
(MTC) and the network is coordinated by Louise
Berg (MTC).
Visit our website, which can be found at KI’s
webpage, under research, research networks:
http://researchnetworks.ki.se/converis/area/183.
Petter Höglund, Chairman
Louise Berg, Coordinator
The KiiM
annual retreat
2006 at
Rosenön.
Department of Microbiology, Tumor and Cell Biology – MTC
79
Emerging Collaborations
The Royal Institute of Technology (KTH)
The interactions between MTC and the
Department of Applied Physics at The Royal
Institute of Technology (KTH) continue to
grow. At the moment, at least three major
collaboration projects are up and running.
KTH provide complementary tools such as
microfabrication techniques, bioimaging,
X-ray, laser technology and spectroscopy, to
the solid biomedical competence at MTC. In
these collaborations novel techniques that
will be useful for future bio-applications are
developed.
Miniaturization of natural killer cell experiments
A research consortium led by the immunologist Petter Höglund (MTC) and including Klas
Kärre (MTC), Björn Önfelt (MTC/Applied
Physics), Karl-Johan Malmberg (Center for
Infectious Medicine, KI) and Ramit Mehr
(Bar-Ilan University, Tel Aviv) study natural
killer (NK) cell tolerance and education. The
project concerns the molecular and cellular
control of NK cell tolerance and education, the
role of MHC class I molecules in this process
and how NK cell education affects the antitumor effects of NK cells. The project includes
several innovative aspects, such as the testing
of a new model explaining NK cell education
in quantitative terms with novel microchipbased imaging experiments developed in the
Önfelt group at Applied Physics, KTH.
Clinical and preclinical approaches will be
fused in a translational effort. In addition, the
team combines biology with biophysics and
mathematics in an interdisciplinary setting.
This research has a potential to lead to define
the optimal conditions for NK cell-mediated
elimination of leukemia and other tumors
using adoptive cell therapy.
Molecular interactions at the single cell
level
Petter Höglund and Jerker Widengren
(Applied Physics) are using fluorescence
correlation spectroscopy (FCS) to study
protein-protein interactions on the surface
of NK cells. FCS is an ultrasensitive spectroscopy technique for studying the mobility of
single molecules. Sofia Johansson, a previous
member of the Höglund laboratory at MTC,
80
Department of Microbiology, Tumor and Cell Biology – MTC
The Royal Institute of Technology (KTH). currently holds a postdoctoral position in the
Widengren laboratory, where she pursues this
project and thus acts as a bridge between the
laboratories.
Novel imaging techniques applied to biological questions
A research project led by Hans Hertz at Applied
Physics is aimed at developing micro imaging
in the x-ray regime. This technique enables
imaging of thick samples in an aqueous environment with high resolution and contrast,
paving the way for biological applications. In
collaboration with Marie Arsenian Henriksson at MTC the technique is being investigated
for its usefulness for small-animal imaging,
especially for tumor detection. In initial studies, mice with defined human neuroblastoma
tumors have been imaged with encouraging
results. The project recently received a collaboration grant founded by the Swedish Research
Council (Vetenskapsrådet).
SciLifeLab Stockholm
Science for Life Laboratory (SciLifeLab)
Stockholm is a national resource center
devoted to large scale bioscientific research.
The center will focus on biomedicine includ-
Photo: courtesy of KTH Photo Archive
ing genomic and proteomic profiling, bioimaging and bioinformatics. SciLifeLab Stockholm was founded through a joined investment by the Karolinska Institutet (KI), KTH
and Stockholm University and will be situated on new premises close to the Karolinska
Campus in Solna.
For more information visit the homepage
http://www.scilifelab.se/. Hjalmar Brismar
at the Department of Applied Physics at
KTH was one of the main applicants and is
currently involved in setting up the bioimaging part of the lab.
Swedish network in bioimaging
The newly formed Swedish BioImaging
Network aims to facilitate collaboration
between researchers in Sweden working in
bioimaging and related fields. Björn Önfelt,
Hans Hertz, Jerker Widengren and Hjalmar
Brismar from the Department of Applied
Physics are members of the network. If you
work in the field of bioimaging and wish
to become a member send an email to [email protected] with a short description of your
bio­imaging activity and interest. Read more
on the homepage http://www.bioimaging.se/.
Björn Önfelt
Emerging Collaborations
Center for Infectious Medicine (CIM)
CIM was established in 2003 by former MTC scientist Hans-Gustaf Ljunggren within the Department of Medicine,
Karolinska University Hospital in Huddinge. The center is actively engaged in immunology and infectious disease research.
In 2007, the groups of Adnane Achour and Benedict Chambers, moved their laboratories to MTC.
Adnane Achour
arrival of my
research group to MTC,
has been extremely positive and resulted in excellent and exciting ongoing scientific collaborations with Birgitta Henriques Normark and
Staffan Normark (on structural studies of
components of the pilus of Streptoccus pneumoniae), Marie Arsenian Henriksson (structural studies of c-Myc), Martin Rottenberg,
Markus Maeurer (development of a tuberculosis vaccine), Mats Wahlgren (studies
of malaria-associated proteins) and Petter
Höglund and Klas Kärre (natural killer cells)
at MTC.
The strategic localization of MTC within
the north campus has also allowed us to
deepen our previous collaborations with
additional research groups at the Karolin-
The
ska University Hospital, Solna such as those
belonging to Håkan Mellstedt and Jacob
Odeberg (development of altered peptide
ligand variants of tumor-associated proteins),
Marie Wahren-Herlenius (modeling of MHC
molecules), Robert A Harris (role of posttranslational modifications in autoimmunity) and Hans Grönlund (crystal structures of
allergens).
Presently, my group is composed of three
PhD students, Adil Doganay Duru, Hannes
Uchtenhagen (main supervisor being AnnaLena Spetz) and Eva Allerbring who will soon
be joined Dr Tatyana Sandalova strengthening our skills and knowledge in structural
biology, especially X-ray crystallography.
Most of our research approach combines
structural biology with biochemistry and
immunology. Our focus has been on the
molecular mechanisms underlying viral
escape from immune recognition and/or the
potential consequences of virus infections,
Active site of the Streptococcus pneumoniaeassociated sortase C (Neiers et al., J Mol Biol 393,
704, 2009).
such as the induction of auto-reactivity. We
have recently initiated a series of cancerrelated studies in which we have developed/
adapted alternative ideas to design altered
peptide ligands. The final aim is to increase
the immunogenicity of wild-type peptides by
introducing mutations that do not alter the
conformation of the targeted peptides.
Benedict Chambers
The focus of my research
is on both dendritic cell
(DC) and natural killer
cell (NK) biology and
how these cells, together
or by themselves, affect
immune responses. DC release a variety of
cytokines and chemokines upon infection
that both attract NK cells to a site of infection and activate these to produce cytokines
or to become cytotoxic. Part of my research is
attempting to identify which DC subsets that
are required for the activation of the NK cells.
One way by which NK cells can control adaptive immune responses is to kill DC. This has to
be avoided in a situation in which DC cells are
used as part of a vaccine strategy. We have identified a variety of molecules on dendritic cells
Live cell imaging (green) of NK-DC cell interactions. Many NK
cells (blue) interact with DC (A). NK cells migrate towards DC
(B), form a tight intercellular contact (immune synapse) with
DC (C) and finally kill DC (D), seen as red staining of the cell
nucleus.
that are involved in NK cell mediated killing
of dendritic cells. These findings are also being
used in developing novel therapies to treat
myeloid derived tumor cells which express
many of the same ligands as dendritic cells.
In viral infections we have found that NK
cells appear to eliminate dendritic cells. The
same was also observed with Toxoplasma
gondii parasite infected DC. Interestingly in
the latter case, the parasite could “jump” into
the NK cells that had killed the dendritic cell.
We are currently investigating how pathogens
such as Toxoplasma, influenza and vaccina
virus interact with NK cells.
We have excellent collaborations with
several research groups at MTC including those that belong to Klas Kärre, Martin
Rottenberg, Gunilla Karlsson Hedestam and
Mikael Rhen.
Department of Microbiology, Tumor and Cell Biology – MTC
81
Translational Research
Successful collaborations between preclinical and clinical scientists
of new drugs and treatment principles to improve human health
requires successful collaborations between
preclinical and clinical scientists. The
modern word for this process is “translational
research”. MTC is primarily a preclinical
department, and as such can provide novel
ideas and biological insights useful for drug
development. In order to stimulate this process, it is important to give good examples on
how it can be successfully conducted.
MTC has several research leaders engaged
in successful translational research. Four such
projects are highlighted here. We have asked
the leaders to describe how the project was
initiated, how they succeed in maintaining it
and how they have overcome potential obstacles that lies in the challenges of communication between preclinical and clinical science.
It is our hope that these short recollections
will inspire the creation of more translational
projects at MTC and other research departments, which will be beneficial for patients in
the future.
The development
Petter Höglund
Anna Nilsson
Name: Yihai Cao
Project title: Vascular endothelial growth
factor (VEGF) in paraneoplastic syndrome.
Collaborators: Dr. Weide Zhaong,
Guangzhou, China.
82
Department of Microbiology, Tumor and Cell Biology – MTC
project was initiated by us when the technological developments made it possible
to immunophenotype T cells during the
reconstitution phase after transplantation. We focus on the behaviour of EBVspecific T cells using tetramer technology
and multicolor flow cytometry.
How is the project maintained?
Name: Ingemar Ernberg, MTC and Markus
Maeurer, MTC and the Swedish Institute of
Infectious Disease Control (SMI).
Project title: Control of Epstein-Barr
virus (EBV) after transplantation, with Special
focus on the Pathogenesis and Control of
EBV-positive lymphomas (SPACE).
Collaborators: Olle Ringdén and
­Brigitta Omazic, (Transplantation Unit, Karolinska University Hospital Huddinge). Jacek
Winiarski and Åsa Gustavsson-Jernberg
(Pediatric clinic, Karolinska University Hospital
Huddinge).
How did you initiate the project and how is it
maintained?
We have had long-standing collaborations with the transplantation unit at
Karolinska University Hospital Huddinge
in projects relating to EBV infection and
stem cell transplantation. The SPACE
We have joint meetings every 5-8 weeks at
the clinic when we go through recent data
and perform a joint analysis. So far, close
to 100 patients have been entered in the
study. They are sampled at 1, 2, 3, 6 and 12
months after transplantation.
Which obstacles have you encountered?
Building a good, open database is difficult
in projects containing clinical data and
patient information.
Which advice do you want to give regarding
the translation of your project?
Build relations. Be humble, show respect
and understand that clinical work with
patients is tedious and needs a good
organization. The lab-work is very much
routine, but still has to be performed at
top level.
How did you initiate the project?
How is the project maintained?
Our animal models had shown that high
levels of VEGF in the circulation globally
affects the vasculature in various tissues
and organs, leading to severe anemia,
hepatomegaly, splenomegaly, and endocrine disorders. Such paraneoplastic
effects are called VEGF-induced cancerassociated systemic syndrome (CASS) and
significantly shortens life time and lowers
quality of life.
We initiated this project to see if circulating levels of VEGF also in human cancer
patients would be reversely correlated with
hematocrit and other cancer-associated
systemic disorders.
I have continuous discussion with the clinicians involved, but we do not have regular
meetings with them.
Which obstacles have you encountered?
Ethical approval of clinical studies is relatively complicated and time consuming.
Which advice do you want to give regarding
the translation of your project?
Start to prepare ethical permission in good
time. Otherwise, it can significantly delay
your studies.
Name: Galina Selivanova
Project title: Rescue of mutant p53 function by the small molecule PRIMA-1.
Collaborators: Klas Wiman (Cancer
Center Karolinska, CCK), Sören Lehmann &
Christer Paul (Department of Hematology, Karolinska Huddinge), Sten Nilsson (Department of
Oncology-Pathology, Karolinska Solna).
How did you initiate the project?
Our preclinical discovery that the activity of mutant p53 could be restored by a
synthetic peptide prompted us to perform
a screen of the chemical library. This led
to the identification of PRIMA-1 as an
apo­ptosis-inducing drug and opened the
door for clinical extensions.
How is the project maintained?
Together with colleagues Klas Wiman and
Staffan Strömblad, and with the help of
Karolinska Development AB, I started a
small biotech company called Aprea AB.
The first clinical trial with PRIMA-1 has
recently started, led by Aprea AB. My group
How did you initiate the project?
Questions formulated in my scientific
group at MTC are proposed to our clinical collaborators. Their experience, clinical
and scientific, then helps to refine the questions and always propose relevant clinical
angles and comments. After the project
is outlined in detail we plan for relevant
samples to be taken from patients provided
that all ethical permissions are in place.
How is the project maintained?
Name: Francesca Chiodi
Project title: B and T cell damage during
HIV-1 infection.
Collaborators: Bo Hejdeman, Sven
Grütz­meier (Stockholm South General Hospital,
Södersjukhuset), Paolo Rossi (Hospital Bambin
Gesu’, Rome Italy), Anna Nilsson (Department
of Woman and Child Health, Karolinska Institutet).
Important components to maintain the
translational collaboration are:
a) reciprocal respect of competence and
culture in the pre-clinical and clinical environments;
b) flow of information and contact during
all stages of the project, including presentation of results to medical staff, both doctors
and nurses;
c) finalize papers with the active participation of clinical collaborators. It is also
important to find common grant support
to give legitimacy to the projects.
Which obstacles have you encountered?
is involved in discussions with Aprea, but
mainly continue with basic research on
the molecular mechanism of action of
PRIMA-1.
Which obstacles have you encountered?
It has been a long road to where we are now.
Along the way, I had to learn many new
things, such as how to communicate with
pharma people, clinicians and chemists.
I also had to learn how to run this
project and at the same time lead the basic
research in my lab.
Which advice do you want to give regarding
the translation of your project?
It is imperative to be dedicated to basic
science and to be passionate about it. But
if something that you have discovered can
help people – put an effort into translating
it into clinic!
Nobody understands the implications
of your discovery better than yourself!
research are, in my experience, not really
between the pre-clinical environment
and the clinic. In my experience the major
obstacle when working with translational
projects in a pre-clinical environment is
often to gain respect from colleagues only
involved in animal experiments.
Also publication in top ranked journals is some time difficult when working
mostly with clinical specimens. But we are
learning to bridge our questions both into
the clinic and in parallel into the animal
house.
Which advice do you want to give regarding
the translation of your project?
If you get an idea for which you need clinical competence and samples, contact clinicians early to plan the project together. You
may get relevant good suggestions which
will increase the value and validity of your
investigations.
Involve clinical collaborators in all
stages of planning and reporting. Never
think about your clinical collaborators as a
sample provider. Be patient if the flow of
specimens to the lab is slow.
The obstacles of conducting translational
Department of Microbiology, Tumor and Cell Biology – MTC
83
Companies Founded by MTC Scientists
Two examples of successful spin-offs
Intervacc AB
The major focus for Intervacc AB is on the
development of vaccines against bacterial
infections within the animal health sector.
Top priority is presently given to a vaccine
against strangles (kvarka in Swedish). This a
very severe and contagious upper respiratory
tract infection in the horse, caused by Streptococcus equi. It leads to severe suffering in the
horse and economic losses for the business.
The world market for a vaccine against strangles is estimated to around 200 million USD.
We have recently proven in a number of
studies on the horse, that a vaccine against S.
equi does not have to be composed of attenuated or killed bacteria, such as those presently
existing. Instead, the use of a well-composed
set of recombinant proteins is both safe and
efficient with no side effects and can meet
the demands from the market for a modern
vaccine.
Intervacc AB is now in a stage where a
proof of principle has been passed and the
candidate vaccine is further optimized, tested
for duration of protection, shelf life, and for
Welsh Mountain Ponies were vaccinated and subsequently challenged with S. equi to assess a protective
effect.
Photo: Jan-Ingmar Flock
the best route of immunisation. Such parameters can be tested directly on the target
animal, the horse, which can be subjected to
experimental infection.
Experience gained from these vaccine
trials is anticipated to facilitate in the development of a vaccine against the human
pathogen Streptococcus pyogenes, having
significant resemblance with S. equi. Several
of the antigens used in the strangles vaccine
are also found in Streptococcus zooepidemicus, another severe pathogen of the horse.
Although there is no cross protection, a
vaccine against S. zooepidemicus is also close
DILAFOR AB
Dilafor AB is a Swedish R&D company
focused on developing pharmaceutical
products from heparin derivatives with low
anticoagulant activity (www.dilafor.com).
12 persons representing a broad scientific-,
drug-development and business expertise
founded the company in 2003 within Karolinska Development AB.
Dilafor AB has a balanced product portfolio representing some highly promising
ideas and innovations. Each of our projects
addresses an important and unfulfilled medical need. All our drug development projects
are based on the use of Low Anticoagulant
Heparin (LAH). Although standard heparin
has numerous biological functions, its potential development as a treatment for indications other than those currently approved is
84
Department of Microbiology, Tumor and Cell Biology – MTC
The bite of an anopheles-mosquito can lead to
malaria.
limited by its anticoagulative properties.
Dilafor has produced several forms of
LAH. Two products are presently in development, tafoxiparin and DF02. Tafoxiparin, which is investigated for its effect on the
prevention and normalization of protracted
labor, has recently completed a “Proof of
Concept” clinical trial in Phase II. Promising results were found in the trial, where 263
women were included, as protracted labor was
prevented in the group given the drug tafoxiparin. Dilafor is also developing a new malaria
at hand and steps have been taken towards
such a vaccine.
The vaccine development also involves a
team at the Swedish University of Agricultural
Sciences, Uppsala and Animal Health Trust
in the UK. Intervacc AB is mainly funded by
the Nordvacc Läkemedel AB and by private
investments. A further spreading of shares is
planned to secure funding for the remaining
parts of the project and for additional developments.
Information about Intervacc AB can be
found at http://intervacc.se/
Jan-Ingmar Flock
treatment – DF02 – that reduces the blocking
of the capillaries in severe Plasmodium falciparum malaria. Today, there are no effective
treatments against the most severe symptoms
of malaria. A Phase I study was conducted
with DF02 during 2009 on 33 healthy men.
The study was randomized, double blind,
and placebo. No severe adverse effects were
reported. The results prove that DF02 is safe
and well tolerated in all of the tested dose
levels both at a single dose and when using
multiple dosing (nine doses during three
days). In addition, the results show that the
drug is suitable for intravenous injection,
which is the only way to treat patients with
the most severe form of malaria. This will be
followed by a proof of concept trial in Phase
II. “New Chemical Entities” similar to tafoxiparin and DF02 are also being developed for
yet other indications.
Mats Wahlgren
Future Faculty
Career development – how to be successful in a large and complex organization
a new seminar series
for the future faculty (postgraduate level)
at MTC was started. The main goals were to
form an interactive group discussing questions such as career development, how to
become a group leader, responsibilities and
challenges, leadership and grant applications
as well as to provide more insights into how to
plan a future career in science.
The first meeting was held in February
2009 in which professor Staffan Normark and
Judit Wefer from the Grant Office at the Karolinska Institutet (KI) discussed important
issues regarding grant applications, where
and how to apply for resources and the evaluation of grant applications. In the next meeting the head of personnel at KI, Ylva Lindberg,
and two human resource consultants, Emma
Liffgarden och Niclas Asplund, talked about
career development and work law.
Three seminars were later held during
fall 2009. The first one was held by the head
During spring 2009
Francoise Barre Sinoussi, who discovered the HIV
virus, has been one of the speakers at the Future
Faculty seminars. Photo: Wikipedia
of MTC, Marie Arsenian Henriksson, who
informed about the organization of MTC
and KI, and discussed the criteria to become a
group leader. The second seminar was held by
the Nobel Laureate in 2008, Francoise BarreSinoussi, who discussed premises on how to
succeed in science with the title “Multidisciplinary science: a key issue in biomedical
research”.
To the last meeting during the fall a representative of both industry and academia was
invited, adjunct Professor Alexander von
Gabain. He was a scientist at KI more than 15
years ago and later co-founded a biotechnology company for development of vaccines,
Intercell Biomedical Research & Development AG, in Austria. He described what it is
like working in industry and how to develop
a company when you come from an academic
background.
Plans for seminars during 2010 include
economic responsibilities and obligations as
a group leader, what is important in leadership and how to be successful in a large and
complex organization like KI.
Birgitta Henriques Normark
Equality and Diversity Group
of the ”Equality and Diversity group” represent scientists, PhD
students, the technical and the administrative personnel within MTC. The main
goal for this group is to promote equal and
respectful treatment irrespective of gender,
age, ethnicity, religion, disabilities or sexual
orientation within all personnel categories. The group is also proactively involved
in different aspects of diversity within the
department.
One important issue that has been
discussed is the role of mentors for the PhD
students. As a result of this discussion, the
mentor function has been more clearly
defined. We have also discussed guide lines
for using English and/or Swedish within the
department.
During the last years, in order to focus
on important issues that cover other
The members
subjects than science, the group has invited
to a number of seminars, that have been
well attended.
In 2007, Stefan Einhorn, professor of
molecular oncology at the Karolinska
Institutet (KI), but also well known as an
author of several popular science books,
spoke about “The art of being kind”. His
lecture was based on a book which has been
a bestseller in Sweden and was translated
into 12 languages. Angela Nilsson from
Stockholm University presented her study
on KI entitled “Innovative or conservative
excellence”.
In 2008, we invited Åsa Björndal and
Laszlo Szekely working at MTC and the
Swedish Institute for Infectious Disease Control, to talk about “Couples in
Research. How to manage a family life in
parallel with hard scientific work”.
We have also
invited Alexander
Perski, who is leading the stress clinic
at KI. He gave a
seminar about “The
Perfection
Trap”.
Hanna Akuffo preAlexander Perski had
an engaged audience
sented her Swedish
in March 2009.
experience
“Just
because you are
paranoid does NOT mean they are not out
to get you - on the other hand perhaps they
are NOT out to get you”. In 2009, David
Eberhard spoke about “Addicted by safety
concerns” and Björn Klinge had a presentation entitled “Ethics”. All these lectures
have been very appreciated and new events
are in the pipe line.
Elisabeth Norin
Department of Microbiology, Tumor and Cell Biology – MTC
85
Undergraduate Education
Teaching in medicine, biomedicine and optometry
in undergraduate education within three study programs; medicine,
biomedicine and optometry, in which we are
responsible for the organization and implementation of courses and are, in addition,
running parts of other courses at the Karolinska Institutet (KI). MTC also harbours, and is
in charge of, one of the two teaching laboratories at Campus Solna, a class II laboratory
equipped to have the capacity for laboratory
work with both bacteria and sterile work with
eukaryotic cells.
Several undergraduate program have
undergone/are undergoing extensive changes
that have required revisions of course contents
and structure. Teachers involved have spent a
lot of time and devotion to form and implement these changes. At MTC there is a continuous effort to further improve the quality of
undergraduate teaching to offer an education
of highest quality.
The medical studies are being reorganized
aiming at a higher level of integration between
disciplines and between preclinical and clinical subjects (LUT = läkarprogrammets nya
utbildningsplan – the New Education Plan for
Studies in Medicine). The former MMI course
(Medical Microbiology and Immunology) at
MTC has been replaced with courses within
DFM1 (den friska människan – the healthy
human being), DSM1 (den sjuka människan
– the sick human being), and DSM2. These
courses contain a mixture of lectures, and to
a large part seminars in small groups, which
have been much appreciated by the students.
The courses consist of both basic immunology,
microbiology and clinical microbiology. The
content of the courses is discussed and evaluated on a regular basis by the course organisers
and the teachers involved to further develop
and maintain the high quality of the courses.
MTC is involved
The Bologna process
With the introduction of the Bologna process
for higher education in Sweden, the study
program in Biomedicine has changed from
being a 4-year program to a 3-year bachelor program and a 2-year Master program
in Biomedicine, starting 2007. MTC is
involved in several courses within the Bachelor program in Biomedicine. At semester 3,
86
Department of Microbiology, Tumor and Cell Biology – MTC
MTC is responsible for giving the course in
“Infection and Immunity” (15 credits). This
course gives the students a basic knowledge in
Immunology and Microbiology and in practical laboratory skills within these subjects.
Teaching is based on lectures, seminars, labo-
ratory work and literature project work. This
course was given for the first time 2008, and
it partly builds on the previous course within
the old 4-year program. It is under constant
evaluation and evolution to maintain high
standards, and student input is an important
part of improving the quality. At semester 5
a course in “Molecular Oncology and Biostatistics”, is given at the department of Oncology and Pathology. MTC is responsible for the
advanced laboratory experiments, a major
and central part of the course which is highly
appreciated by the students.
The Master program in Biomedicine is a
2-year international study program attracting students from all over the world. During
the first semester, the course in “Frontiers
in Translational Medicine” (22 credits) is
intended to introduce the students to some
major research areas at Karolinska Institutet.
The course consists of four parts, MTC is in
charge of two of them; Infection biology and
Oncology. In collaboration with the Center
for Infectious Medicine (CIM), Department
of Medicine in Huddinge, MTC also takes
part in teaching “Immunology and Infection biology” during the second year of the
Master program in Biomedicine. Teaching
is performed in close collaboration with the
PhD courses given at MTC and CIM, and
is an excellent opportunity to attract top
students for future doctoral studies to MTC.
Bachelor and Master programs
MTC is also responsible for project courses
and degree projects for students at the Bachelor and Master programs within Biomedicine and for incoming exchange students on
these programs. The students do a research
project with a research group at KI, any other
university in the world, or at a company, and
at the end of the course present this work
orally and in a written report.
The study programs in Biomedicine have
introduced the 7 grade system A-F, according
to the Bologna process. A lot of work is put
into the development of how students are
examined and graded in these programs and
how to formulate course goals and grading
criteria that can be assessed in 7 grades.
In addition MTC is involved in the Study
program in Optometry in which the students
get tailor made lectures to cover important
aspects of microbiology in this area for opticians.
Jonas Sundbäck
Jan-Ingmar Flock
Thi Viet Ha Nguyen from Vietnam defends her thesis “Diagnosis and treatment of Helicobacter
pylori infection in Vietnamese children” November 2009.
Training for the Future – PhD Studies
The third level of education, formerly
designed as doctoral or postgraduate
education, is very vivid at MTC, with
about 150 students from 30 different
countries guided by more than 65 supervisors. As usual at the Karolinska Institutet (KI), the main education takes place
as scientific work in individual research
groups under the guidance of at least two
supervisors.
There are several “sub-subjects” to
register in, such as infection biology,
immunobiology, cell biology and tumor
biology, biomedical ecology and infectious disease control. The administration is handed by a full time employed
administrator and a study counselor
who, together with the Committee for
Doctoral Education and the MTC leadership, assure that all regulations issued
by KI are followed during registration
for post-doctoral education, half time
control and at the dissertation process.
MTC has two special rules. All students
give in connection with their yearly follow
up, a 10–15 min seminar, and get questions from the audience and feedback
from an independent senior scientist. An
oral examination in the basic knowledge
in microbiology and immunology or in
cell and tumor biology is performed for
each student by an independent senior
scientist before the half time control.
During the last years around 30 new
students have registered per year and the
same number have passed their dissertation. Two thirds are women, whereby
among the Swedish students only one
out of five is male (see Figure). This is an
unusual gender imbalance compared to
KI as a whole.
Females
foreign 25%
Males
foreign 27%
Females
Swedish
39%
Males
Swedish 9%
The median age is 30 years, which is
younger than for the average at KI and the
median time from registration to dissertation is 4.6 years. The median age at dissertation is lower than that for KI total. 18
students (12 %) have a medical exam, half
of which are from Sweden. In total only 18
students have had their basic education at
KI, which shows the need for highly active
external recruitment.
A variety of scientific seminars, usually
one per day, are offered at MTC. Furthermore, many research groups arrange their
own seminar series. In addition, in 2009,
MTC was involved in 15 postgraduate
courses arranged by KI.
Roland Möllby
Department of Microbiology, Tumor and Cell Biology – MTC
87
Summer Research
Schools – a fantastic opportunity
for high school and
undergraduate
students to get a
research experience at KI and
MTC.
Summer Research Schools
An excellent opportunity for students to get a research experience
three summer research
schools at the Karolinska Institutet (KI):
The Summer Research School in Biomedical
Sciences for Swedish high school students,
The Summer Course in Medical Research for
KI undergraduate students and The Amgen
Scholars Program for undergraduate students
from all over Europe.
The Summer Research School in Biomedical Sciences is open for all high school
students that study either natural science or
technical programs in Sweden. The interest
is very high and 20 students are accepted each
summer. Students from outside of Stockholm
are offered a student room during their stay.
The school is five weeks long and consists
of lectures, study visits to a life science
company and to the Nobel museum and a
4-week research project in a research group
at KI. Many research groups at MTC hosted
these students. There is also a social program
for the students including a number of activities during the summer which is organized by
undergraduate students from the KI.
The school ends with student presenta-
MTC administrates
88
Department of Microbiology, Tumor and Cell Biology – MTC
tions and a farewell dinner. Many of these
former students are now undergraduate
students at the KI, with a strong interest in
research!
The Summer Course in Medical Research
(10.5 credit points) is open only for undergraduate students at KI, to get research experience in a research group. 80-90 students from
many of our study programs at the KI apply
for the course which accepts 38 students. It
starts in mid June with lectures in ethics, reading of research publications, learning to plan
and create posters and powerpoint presentations and an inspirational lecture about creativity by Ingemar Ernberg, professor at MTC.
The students take part in a six week project
during the summer in a research group at KI,
many at MTC, after which they present their
results in a written report, a poster and an oral
presentation during two days in August.
The Amgen Scholars Program is new
to Europe and KI since the summer of 2009.
It is supported by the Amgen Foundation
at Amgen, a major biotechnology company
located in the US. Similar sister programs
are located at the University of Cambridge,
UK, and Ludwig-Maximilians University in
Munich, Germany. It is an international summer research school for top undergraduate
students from all over Europe, as defined by
the Bologna process. In the first year 12 students where admitted. During the eight weeks
of the program, the students got an introduction to research, how to present data, make
posters and publish papers. The students also
attended a few research presentations. Most
of the time, however, was spent in research
laboratories all over KI, including MTC. The
program ended by student presentations and a
final symposium at the Cambridge University
which included all Amgen Scholars Programs
in Europe in which the students presented their
projects on posters.
In summary, there is a fantastic opportunity
for students to get a top-level research experience at the KI and MTC and vice versa an excellent opportunity for our research groups to get
in contact with students who are highly interested in research and in future doctoral studies.
Jonas Sundbäck
Life as a PhD Student
One of your greatest adventures!
ing, supervision, language skills, teaching and
presentation techniques.
One of the main obstacles that I have
found hardest to overcome is keeping the
motivation and to continue working hard
even though useful results can be absent for
long periods of time. Since PhD students in
a way are their own managers, their motivation is the only fuel that can run the ongoing projects. During those periods when the
motivation has been failing me it has been
very helpful to find it again elsewhere than
in the lab, e.g. by seeking support from the
supervisor, my mentor, colleagues, group
members and from social activities. It is also
To become a PhD student
always good to once in a while remember the
Supervision is crucial in the beginning since
ultimate goal of the research project and the
most PhD students are beginners within
importance of it in a bigger picture, and not
their fields! Finding a good supervisor that
just as a cell culture in
I got along with well
your Petri dish.
was one of my highWorking
with
est priorities when
science in a research
I started thinking
group might involve
about becoming a
very intense periods
PhD student. Eventuwith a lot of work, but
ally, everything finally
also freedom. A freefelt good at MTC in
dom that gives you the
every way! One could
luxury to decide over
say that the interestyour own work and
ing research projects,
your own time. The
the highly compePhD student period
tent supervision and
has the capacity of
people, the high qualbecoming one of the
ity of the laboratories,
Malin Stoltz performing an experiment.
greatest adventures of
the nice premises,
your entire professional career.
and the pleasant and stimulating research
atmosphere all made me feel welcome and
motivated to stay. Now when the dissertation
The future
is approaching rapidly I have not regretted
The end of my PhD studies is approaching
becoming a PhD student at MTC even once.
and the question is: what will happen next?
Knowing that the dissertation will come
It feels like a PhD degree never can be wrong
soon is of course a bit stressful, but a natural
when applying for positions in the future.
part of the process of being a PhD student.
I have learned so much, and my whole way
The path towards the dissertation is not
of thinking has been totally changed during
straight and is for sure full of hard work, hopethese years, even though I have not started
fully giving the PhD student the opportunity
writing the thesis yet! It seems to me that this
to mature and gain more and more specific
education has taken me to a point where I can
knowledge and independence during the way.
become whatever I want.
In the end, except from the title, becoming a
These years have given me a very solid
PhD also involves personal development and
ground to stand on during the rest of my
achieving skills such as project leading, labocareer and I am grateful for that.
ratory techniques, communication, networkMalin Stoltz
Before writing this I counted backwards
and realized that I have been working in the
same group at MTC/the Swedish Institute of
Infectious Disease Control (SMI) for almost
five years and that I still like it! During 20042005, I attended Stockholm Research School
in Molecular Life Sciences and did one of my
three 10-week projects at MTC/SMI. Before
this I was sceptical about conducting PhD
studies and convinced that I would start in a
pharmaceutical company. However, working in a highly stimulating research milieu at
MTC/SMI made me change my mind!
The MTC PUB –
a lot of fun every month!
One Friday every month there is a
special night at MTC, a night when
the party goes on until 2:00 or 3:00
in the morning – depending on how
long the beers and cocktails last!
Traditionally, MTC has one of the
most appreciated pubs on the Solna
campus and is actually considered
one of the best party evenings you can
find! The very experienced “Muppet
Crew”, which ran more than 30 pubs,
recently left the responsibility to
organize this event in the hands of
the grand “Viceversas Crew”. It was a
challenge for the Viceversas to maintain the high level of quality and fun
that the Muppet Crew had created.
They were able to transform a normal
pub into a real party, where they
allowed everyone to travel, with their
thematic pubs, all over the world,
exploring the cuisine of every possible country...
Bringing a new twist
to the pub evenings,
the Viceversas
are doing a
superb job
reinterpreting
movie themes,
dressing in
“particular”,
eccentric ways
and overall playing good music and
preparing high quality
sandwiches. It is clear that the trend
remains; people prefer the MTC pubs
to those that you find in the downtown area!
Simone Pensioroso
Department of Microbiology, Tumor and Cell Biology – MTC
89
Doctoral Theses 2007–2009
2007
297. Sandalova, Elena,
Regulation of the proapoptotic protein Bim by T cell receptor triggering in human T cells. Opponent: Philippe
Bouillet, Parkville, Australia. Supervisor:
Victor Levitsky.
The role of IL-7 in
Lymphopenia and Bystander Apoptosis
during HIV-1 infection. Opponent: MarieLise Gougeon, Paris, France. Supervisor:
Francesca Chiodi.
298. Fluur, Caroline,
Visualizing the dynamic
interplay between the host and bacterial
pathogens. A real-time study of renal infection. Opponent: Michael S Donnenberg,
Baltimore, USA. Supervisor: Agneta RichterDahlfors.
305. Månsson, Lisa,
306. Hidmark, Åsa, Induction of type I interferons and viral immunity. Opponent: Bruce
Beutler, La Jolla, USA. Supervisors: Gunilla
Karlsson-Hedestam, Eva Nordström, Peter
Liljeström.
Carbapenem resistance in Pseudomonas aeruginosa. Opponent:
Giuseppe Cornaglia, Verona, Italy. Supervisors: Bengt Wretlind, Göran Kronvall.
307. Corrigan, Gary, Evaluation of reverse
transcriptase assay for viral load monitoring.
Opponent: Elizabeth M Dax, Fitzroy, Victoria,
Australia. Supervisors: Rigmor Thorstensson,
Clas Källander.
300. Advani, Abdolreza, Epidemiological characterisation of Bordetella pertussis in Sweden,
1970–2004. Opponent: Per Olcén, Örebro,
Sweden. Supervisors: Hans Hallander, Lars
Engstrand, Patrick Olin.
308. Paulander, Wilhelm, Mechanisms of adaptation to the fitness cost of antibiotic resistance. Opponent: Ivan Matic, Paris, France.
Supervisors: Dan Andersson, Sophie Maisnier-Patin.
299. Giske, Christian G,
Genomic and transcriptomic
variation in blood stage Plasmodium falciparum. Opponent: Hernando del Portillo,
Barcelona, Spain. Supervisor: Mats Wahlgren.
309. Mok, Bobo,
Regulation and characterization of antimicrobial peptides in man
and mice. Opponent: Peter Hiemstra, Leiden,
The Netherlands. Supervisors: Mats Andersson, Katrin Pütsep.
310. Karlsson, Jenny,
311. Bråve, Andreas, Prime-boost immunization strategies against HIV-1. Opponent:
Tomás Hanke, Oxford, UK. Supervisors:
Britta Wahren, Karl Ljungberg, Margaret Liu.
Studies of cell signaling
using bacterial toxins and organic electronic
devices. Opponent: Thomas Borén, Umeå,
Sweden. Supervisor: Agneta Richter-Dahlfors.
312. Kjäll, Peter,
301. Sjöström, Karin, Molecular epidemiology of pneumococcal carriage and invasive
disease. Opponent: Karl Kristinsson, Reykjavik, Iceland. Supervisor: Birgitta Henriques
Normark.
Intercellular protein
transfer and regulation of inhibitory NK
receptor accessibility. Opponent: Francesco
Colucci, Cambridge, UK. Supervisors: Petter
Höglund, Klas Kärre, Maria Johansson.
302. Andersson, Katja,
303. Ries, Johannes, Pneumococcal pili and
other cell surface properties affect the infection biology of Streptococcus pneumoniae.
Opponent: Regine Hakenbeck, Kaiserslauten,
Germany. Supervisors: Birgitta Henriques
Normark, Staffan Normark, Eva Morfeldt.
304. Darai-Ramqvist, Eva, Involvement of
evolutionarily plastic regions in cancer associated chromosome 3 aberrations. Opponent:
David Gisselson, Lund, Sweden. Supervisors:
Stefan Imreh, Maria Kost-Alimova.
PhD student Xuan Xue follows the old tradition of nailing his thesis with a golden nail.
90
Department of Microbiology, Tumor and Cell Biology – MTC
EBV gene variation and
epigenetic alterations in Asian nasopharyngeal carcinoma and potential clinical applications. Opponent: Jaap M Middeldorp,
Amsterdam, The Netherlands. Supervisors:
LiFu Hu, Ingemar Ernberg, Phan Thi Phi Phi,
Tran Thi Chinh.
313. Nguyen-Van, Do,
314. Liu, Anquan, Proinflammatory factor
mediated lymphocyte activation – the pivotal
role of leukotriene B4. Opponent: Dorothy H
Crawford, Edinburgh, UK. Supervisors: Eva
Klein, Laszlo Szekely.
Chemokines and chemo­
kine receptors during viral infections in man.
Opponent: John Gordon, Birmingham, UK.
Supervisors: Anna Nilsson, Francesca Chiodi.
315. Mowafi, Frida,
Characterization of c-diGMP signaling in Salmonella typhimurium.
Opponent: Stephen Lory, Boston, USA.
Supervisors: Ute Römling, Mats Andersson.
316. Simm, Roger,
317. Vahlne, Gustaf, Natural killer cell inhibitory and activating receptors – Regulatory
role in effector functions against normal and
tumor cells. Opponent: John Torgils Vaage,
Copenhagen, Denmark. Supervisors: Maria
Johansson, Klas Kärre, Petter Höglund.
Modulation of the deubiquitinating system in viral infection, lymphoid
cell activation and malignant transformation.
Opponent: Ivan Dikic, Frankfurt, Germany.
Supervisors: Maria G Masucci, Riccardo
Gavioli.
318. Rolén, Ulrika,
319. van der Holst, Rutger, TRAPC a novel
Triggering Receptor expressed on Antigen
Presenting Cells. Opponent: Marco Colonna,
St Louis, USA. Supervisors: Jonas Sundbäck,
Klas Kärre.
320. Fernebro, Jenny, Genetic approaches
towards
understanding
pneumococcal
virulence and biology. Opponent: Tone
Tonjum, Oslo, Norway. Supervisors: Birgitta
Henriques Normark, Staffan Normark.
2008
Identification and characterization of novel virulence factors in
Streptococcus pneumoniae. Opponent: Sven
Hammerschmidt, Munich, Germany. Supervisors: Birgitta Henriques Normark, Staffan
Normark.
321. Wartha, Florian,
The Head Marshall Ying Zhao (foreground) from MTC leads the opening of the KI Conferment Ceremony
May 2009. Photo:Ulf Sirborn
322. Beiter, Katharina, Recognition and clearance of Streptococcus pneumoniae by the
innate immune system. Opponent: Malak
Kolb, Nashville, USA. Supervisors: Birgitta
Henriques Normark, Staffan Normark.
328. Tallo, Tatjana, Molecular epidemiology of
323. Svanh, Anita, Varicella immunity and
vaccination. Opponent: Tomas Bergström,
Gothenburg, Sweden. Supervisors: Annika
Linde, Hans Gaines.
329. Reiner, David,
324. Meyer-Hoffert, Ulf, Antimicrobial peptides
as defense molecules at the interface of
the host and bacteria. Opponent: Artur
Schmidtchen, Lund, Sweden. Supervisors:
Birgitta Henriques Normark, Jens-Michael
Schröder, Staffan Normark.
330. Andersson, Ida, Crimean-Congo hemorrhagic fever virus: Interferon-induced antiviral mechanisms and immune evasion strategies. Opponent: Richard Elliott, St. Andrews,
UK. Supervisors: Ali Mirazami, Åke Lundkvist.
325. Juréen, Pontus,
Molecular characterisation of antibiotic resistance in Mycobacterium
tuberculosis. Opponent: Viesturs Baumanis,
Riga, Latvia. Supervisors: Sven E Hoffner, Lars
Engstrand.
331. Skoglund, Anna,
326. Kader, Mohammad Abdul, GGDEF-EAL
domain proteins in bacterial physiology.
Opponent: Iñigo Lasa Uzcudun, Navarra,
Spain. Supervisors: Ute Römling, Mats Wahlgren.
332. Maljkovic Berry, Irina,
hepatitis A, B and C in Estonia. Opponent:
Magnus Lindh, Gothenburg, Sweden. Supervisors: Helene Norder, Lars Magnius, Ljudmilla Priimägi, Valentina Tefanova.
Cell cycle and differentiation in Giardia lamblia. Opponent: Norbert
Müller, Bern, Switzerland. Supervisors:
Staffan Svärd, Frances Gillin.
Mechanisms of genetic
adaptation in Helicobacter pylori. Opponent:
Arnoud van Vliet, Norwich, UK. Supervisors:
Lars Engstrand, Margareta Krabbe och Britta
Björkholm.
Genetic aspects of
HIV-1 evolution and transmission. Opponent; Andrew Leigh Brown, Edinburgh, UK.
Supervisors:Thomas Leitner, Jan Albert.
Pursuing the fever trail.
Pathogenesis of blood-stage P. falciparum
malaria & pregnancy. Opponent: Ross
Coppel, Victoria, Australia. Supervisors: Mats
Wahlgren, Qijun Chen, Craig Wheelock.
333. Rasti, Niloofar,
Immunological consequences of Epstein-Barr virus
replication. Opponent: Emmanuel Wiertz,
Leiden, The Netherlands. Supervisors: Victor
Levitsky, Maria Masucci.
327. Ortlieb Guerreiro Cacais, André,
Department of Microbiology, Tumor and Cell Biology – MTC
91
334. Hardestam, Jonas, Hantavirus – shedding,
stability and induction of apoptosis. Opponent: Göran Wadell, Umeå, Sweden. Supervisors: Åke Lundkvist, Jonas Klingström.
335. Aronsson, Linda, Microbial regulation
of PPAR: Nuclear receptor networks important for colonic homeostasis. Opponent:
Ian Sanderson, London, UK. Supervisors:
Velmurugesan Arulampalam, Sven Pettersson, Jan-Åke Gustafsson.
336. Rölle, Alexander, Natural killer cell mediated recognition of herpesviruses – Mechanisms of viral immune escape. Opponent:
Andrew McMichael, Oxford, UK. Supervisors: Klas Kärre, Cristina Cerboni, Adnane
Achour.
337. Forsell, Mattias, Evaluation of new strategies to stimulate neutralizing antibodies
against HIV-1. Opponent: Quentin Sattentau,
Oxford, UK. Supervisors: Gunilla Karlsson
Hedestam, Richard Wyatt.
338. Åkerström, Sara, SARS coronavirus – The
role of occessory proteins and nitric oxide
in the replication cycle. Opponent: Völker
Thiel, St Gallen, Switzerland. Supervisors: Ali
Mirazimi, Yee-Joo Tan, Åke Lundkvist.
339. Blomberg, Christel, Insight into the genetic
characteristics of pneumococcal isolates.
Opponent: Peter Hermans, Nijmegen, The
Netherlands. Supervisors: Birgitta Henriques
Normark, Eva Morfeldt, Staffan Normark.
Innate immunity to
intracellular bacterial infections. Opponent:
Thomas Decker, Vienna, Austria. Supervisors:
Martin Rottenberg, Hans Wigzell.
Källman, Owe, Antibiotic resistance
in Gram-negative bacilli – with focus on
cephalosporin resistance mechanisms in
Escherichia coli and Klebsiella pneumoniae.
Opponent: Niels Frimodt-Möller, Copenhagen, Denmark. Supervisors: Barbro OlssonLiljequist, Christian G Giske, Mats Kalin.
341.
342. Simon, Melinda, Crimean-Congo hemorrhagic fever virus: Interactions with host cell
structures in viral replications. Opponent:
Einar Everitt, Lund, Sweden. Supervisors: Ali
Mirazimi, Åke Lundkvist.
Transcriptome based
analysis of Hodgkin lymphoma: Insights into
the microenvironment. Opponent: Volker
Diehl, Cologne, Germany. Supervisor: Ingemar Ernberg.
343. Birgersdotter, Anna,
344. Tran, Thi Thanh Ha, HIV-1 genotype and
vertical transmission in Northern Vietnam.
Opponent: Birgitta Åsjö, Bergen, Norway.
Supervisors: Francesca Chiodi, Phung Dac
Cam, Anneka Ehrnst, Anna Nilsson.
345. Bucardo, Filemon, Pediatric rotavirus and
norovirus diarrhea in Nicaragua. Opponent:
Tomas Bergström, Gothenburg, Sweden.
Supervisors: Lennart Svensson, Roland
Möllby, Felix Espinoza.
346. Wang, Xiaoda, Characterization of Esche­
richia coli colonizing the gastrointestinal tract
and urinary tract catheters. Opponent: Timo
Korhonen, Helsinki, Finland. Supervisors:
Ute Römling, Annelie Brauner.
340. Trumstedt, Christian,
Surface antigens and
virulence in Plasmodium falciparum malaria.
Opponent: Alister Craig, Liverpool, UK.
Supervisors: Mats Wahlgren, Gerhard Winter.
348. Rotzén Östlund, Maria, Viral respiratory
infections: Diagnosis and Epidemiology.
Opponent: Anders Widell, Lund, Sweden.
Supervisors: Lena Grillner, Tobias Allander,
Benita Zweygberg Wirgart.
Molecular characterization of Mycobacterium tuberculosis complex
from Kampala, Uganda. Opponent: Sebastien
Gagneux, London, UK. Supervisors: Gunilla
Källenius, Moses Jolaba, Tuija Koivula.
349. Asiimwe, Benon,
2009
350. Cagigi, Alberto, HIV-1 infection and loss
of serological memory: the role of altered
expression of B-cell chemokine receptors,
timing of HAART and impaired antibody
affinity maturation. Opponent: Rita Carsetti,
Rome, Italy. Supervisors: Anna Nilsson,
Francesca Chiodi.
351. Viklund, Ing-Marie, Identification and
characterization of WASP and FKBP-like
protein. Opponent: Maria Fällman, Umeå,
Sweden. Supervisors: Sven Pettersson, Pontus
Aspenström.
Vaccination against
drug-resistant HIV. Opponent: Fabrizio
Manca, Genoa, Italy. Supervisors: Maria
Isaguliants, Britta Wahren, Annika Karlsson.
352. Boberg, Andreas,
353. Ridderstråle, Karin, Myc oncoproteincofactor interactions; function, regulation
and targeting. Opponent: Jonas Nilsson,
Umeå, Sweden. Supervisors: Lars-Gunnar
Larsson, Ola Söderberg, Natalie von der Lehr.
347. Normark, Johan,
354. Chêne, Arnaud, Impact of malaria on B-cell
homeostasis and Epstein-Barr virus reactivation. Endemic Burkitt´s lymphoma pathogenesis. Opponent: Maria Manuel Mota, Lisbon,
Portugal. Supervisors: Mats Wahlgren, Maria
Teresa Bejarano, Adnane Achour.
Signaling pathways controlling bacterial adaption. Opponent: Dieter
Haas, Lausanne, Switzerland. Supervisors:
Öjar Melefors, Ute Römling.
355. Jonas, Kristina,
356. Vudattu, Nalini Kumar, Alternative splicing
of interleukin-7. Opponent: Rhodri Ceredig,
Ireland. Supervisors: Markus Maeurer, Maria
Masucci.
Jenny Sjöberg from MTC receives her Doctoral hat at the Karolinska Institutet Conferment Ceremony
Photo: Ulf Sirborn
May 8th 2008. 92
Department of Microbiology, Tumor and Cell Biology – MTC
Studies on regulatory
networks governing virulence gene transcription in Staphylococcus aureus, Opponent: Paul
Williams, Nottingham, UK. Supervisors: Patric
Nilsson, Staffan Arvidson, Rune Andersson.
357. Gustafsson, Erik,
Innate and adaptive
immune responses in viral and chronic
inflammatory diseases. Opponent: Vincenzo
Banaba, Rome, Italy. Supervisors: Louise
Berg, Klas Kärre.
358. Schepis, Danika,
359. Magalhaes, Isabelle, CD8α+ T cells and
immune memory. Opponent: Pedro Romero,
Lausanne, Switzerland. Supervisors: Markus
Maeurer, Rigmor Thorstensson.
Amoebiasis: Diagnosis and
prevalence in León Nicaragua. Opponent:
Agneta Aust-Kettis, Uppsala, Sweden. Supervisors: Ewert Linder, Jadwiga Winiecka-Krusnell, Roland Möllby.
360. Leiva, Byron,
Dissecting the p53 pathway
by means of small molecule-mediated reactivation and computational biology. Opponent: Michael Resnick, Research Triangle
Park, USA. Supervisor: Galina Selivanova.
361. Enge, Martin,
362. Vivar, Nancy, Altered T cell homeostasis
during HIV-1 infection: Consequences of
lymphopenia and chronic T cell activation.
Opponent: Franco Lori, Washington, USA.
Supervisors: Bence Rethi, Francesca Chiodi.
Gut microflora associated
characteristics in children with celiac disease.
Opponent: Knut Lundin, Oslo, Norway.
Supervisors: Elisabeth Norin, Tore Midtvedt,
Lars Stenhammar.
363. Tjellström, Bo,
Molecular mechanisms of growth suppression by pharmacologically activated p53. Opponent: Jean-Christoph Marine, Gent, Belgium. Supervisors:
Galina Selivanova, Joanna Zawacka-Pankau.
364. Hedström, Elisabeth,
365. Mphande, Fingani Annie, Towards understanding the SURFIN protein family and var
genes in Plasmodium falciparum. Opponent:
Jürgen Kun, Tubingen, Germany. Supervisors: Mats Wahlgren, Fred Kironde, Gerhard
Winter.
366. Kis, Loránd Levente, The role of the micro-
environment on the regulation of EpsteinBarr virus latent gene expression. Opponent:
Lawrence S. Young, Birmingham, UK. Supervisors: Eva Klein, Laszlo Szekely.
Display of recently finished doctoral thesis at the entrance of MTC.
367. Huitric, Emma, Drug resistance in M.
tuberculosis and the characterization of a new
anti-tuberculosis drug candidate. Opponent:
Stewart Cole, Lausanne, Switzerland. Supervisors: Sven Hoffner, Dan Andersson, Koen
Andries.
368. Gudmundsdotter, Lindvi, HIV-1 immune
responses induced by natural infection or
immunisation. Opponent: Harriet Robinson,
Atlanta, USA. Supervisors: Britta Wahren,
Frances Gotch, Nesrina Imami, Rigmor
Thorstensson.
369. Benno, Peter, Biochemical parameters
reflecting the intestinal ecology of healthy
adults and alterations of these parameters in
patients with ulcerative colitis or rheumatoid
arthritis. Opponent: Per Falk, Princeton, USA.
Supervisors: Tore Midtvedt, Elisabeth Norin.
370. Näslund Tanja, Viral vectors in innate
and adaptive immunity. Opponent: Gerd
Sutter, Munich, Germany. Supervisors: Peter
Liljeström, Margaret Chen.
371. Negrea, Aurel, Pharmaceutical
and mutational interference with virulence of Salmonella enterica serovar Typhimurium. Opponent: Josep Casadesus, Seville, Spain. Supervisor: Mikael Rhen.
372. Ribacke, Ulf, Parasite virulence and disease
severity in Plasmodium falciparum malaria.
Opponent: Manuel Llinás, Prince­ton, USA.
Supervisors: Mats Wahlgren, Peter Nilsson.
373. Dellacasa Lindberg, Isabel, Dissemination of Toxoplasma gondii to the central
nervous system – with special reference to in
vivo bioluminescence imaging. Opponent:
Geneviève Milon, Paris, France. Supervisors:
Antonio Barragan, Mats Wahlgren.
374. Kilewo, Charles, Prevention of mother-to-
child transmission of HIV-1 using antiretroviral drugs in Dar es Salaam, Tanzania. Oppo-
nent: Thorkild Tylleskär, Bergen, Norway.
Supervisors: Gunnel Biberfeld, Fred Mhalu,
Andrew Swai.
375. Gaseitsiwe, Simani, Infections with HIV-1
and Mycobacterium tuberculosis; the Role of
HLA Class II alleles and HIV-1 phenotypes.
Opponent: Sven Britton, Solna, Sweden.
Supervisors: Anneka Ehrnst, Markus Meurer,
Andrzej Pawlowski, Max Essex.
Genetic adaptions of Helicobacter pylori during gastric
disease progression. Opponent: Stefan
Bereswill, Berlin, Germany. Supervisors: Lars
Engstrand, Britta Björkholm.
376. Kling Bäckhed, Helene,
377. Xue, Yuan, Mechanism of pathological
angiogenesis in adipose tissue and tumor.
Opponent: Paul Trayhurn, Liverpool, UK.
Supervisor: Yihai Cao.
Genetic content of
clinical pneumococcal isolates and its relation to disease outcome. Opponent: Pentti
Huovinen, Åbo, Finland. Supervisor: Birgitta
Henriques Normark, Eva Morfeldt.
378. Dagerhamn, Jessica,
Characterization of host
and microbe interactions affect, adherence,
clearance and systemic spread of S. pneumoniae. Opponent: Ingileif Jónsdóttir, Reykjavik, Iceland. Supervisors: Birgitta Henriques
Normark, Staffan Normark, Barbara Albiger.
379. Dahlberg, Sofia,
380. Muschiol Sandra, Small molecule inhibitors of type III secretion and their effect
on Chlamydia development. Opponent:
Kenneth Fields, Miami, USA. Supervisors: Birgitta Henriques Normark, Staffan
Normark, Agathe Subtil.
381. Nguyen, Thi Viet Ha, Diagnosis and treatment of Helicobacter pylori infection in Vietnamese children. Opponent: Peter Unge,
Örebro, Sweden. Supervisors: Marta Granström, Gia Khanh Nguyen, Mikael Sörberg.
Department of Microbiology, Tumor and Cell Biology – MTC
93
Core Facilities
MTC FACS Facility
To the forefront of flow cytometry technology
core facility at MTC
originates from the Fluorescence Activated
Cell Sorting (FACS) unit at the former Tumor
Biology Department established in 1983. It is
thus the oldest core facility for flow cytometry
in the Stockholm area.
During recent years flow cytometry technology has developed rapidly. Instruments are
now able to analyze large numbers of different
fluorochromes simultaneously and in parallel there has been an expansion in the number
of fluorescent dyes, directly conjugated antibodies and other reagents. This allows more
advanced and accurate analysis of lymphocyte
as well as other cell populations.
MTC has during 2008–2009 moved to
the forefront of flow cytometry technology
through the support of the Knut and Alice
Wallenberg foundation. We have obtained
three new instruments. One advanced 11
color MoFlo XDP cell sorter which allows
high-speed, four-way sorting, an LSR II cell
analyzer with 18 color detection and a CyAn
ADP instrument for 7 color analysis.
The flow cytometry
The FACS Facility at MTC has customers from more than 50 research groups at MTC and KI as well as external
customers. Birgitta Wester operating the LSR-II flow cytometer, while a user is preparing samples.
We have also obtained new computers and
software for data analysis. To accommodate
these new instruments we have expanded the
facility. Altogether we now offer two instruments for cell sorting, five instruments for cell
analysis and three computers for data analysis.
We are now in the process of upgrading our
CyAn ADP to 9 colors and equipping it with a
plate-loader for automated analysis of samples
in 96 well plates.
Multicolor flow cytometry allows
phenotypic and functional analysis of rare
subpopulations of lymphocytes, it is thus
possible to identify over 100 subpopulations
in human blood. One may also study production of cytokines and degranulation via cell
surface-mobilization of a marker for secretory lysosomes in small subpopulations of
lymphocytes.
The figure shows an example of functional
analysis of subsets of mouse natural killer
(NK) cells.
Maria Johansson
A 12-color flow cytometric analysis of cytokines (Mip1a and IFNg) and degranulation (CD107a) on subsets of mouse NK cells upon stimulation of the activating receptor NKp46. Courtesy Petter Brodin.
94
Department of Microbiology, Tumor and Cell Biology – MTC
Birgitta Wester
Mikael Jondal
Core Facilities
KI Visualization Facility (KIVIF)
Providing researchers with frontline imaging technology
Modern imaging technology finally made
it possible to directly observe some of the
most intricate details of the living material.
Today we can visualize such delicate events
as the movements of individual macromolecules or different cellular organelles in live
cells or the subtle interplay between microbes
and their host cells. With the right equipment
and approach, biological imaging has become
not only an illustrative tool but also a scientific method of its own.
KIVIF at MTC was established in January 2004 and was officially inaugurated in
November 2005 by KI President Harriet Wallberg Henriksson.
The goal of KIVIF is to provide researchers
at the Karolinska Institutet (KI) with frontline imaging technology in combination with
technical support and imaging-related expert
advice within the frame of their own scientific
projects. The facility has around two hundred
active users every year, mainly from within
KI, but some researcher also come from other
universities.
KIVIF offers a broad range of imaging
technologies; from basic fluorescence and
phase contrast microscopy to advanced laser
confocal live cell imaging. KIVIF also provides
assistance with routine histology and sample
preparation.
The spinning disc confocal microscope in use at the KI Visualization Facility at MTC.
Confocal microscopes
At present the facility has the following instruments:
n Three confocal microscopes:
– a Perkin Elmer ERS system with microlens enhanced Nipkow spinning disc and
5 laser line illumination, equipped with a
complementary laser engine for pixel precise
burning capacity for FRAP, FLIP, acceptor
bleaching FRET and photoconversion experiments.
– a fully automated Perkin
Elmer LCI spinning disc system
with programmable XY stage
and integrated image capture
and on-the-fly image analysis capacity for extended field
confocal microscopy, massively
parallel filming and high
throughput imaging.
– a newly obtained Leica TCS
SP5 X laser scanning confocal
system with a continuous spectrum white laser for imaging of
every possible fluorochromes
and recombinant fluorescent
proteins.
n Three conventional fluoresConfocal live cell imaging of Human prostate tumor cells (PC3);
cence
microscope with cold CCD
Z-stack projection at one timepoint. Green: b-actin (cytoskeleton), red: Histone 2A/m Nuclei.
cameras for routine imaging.
n An atomic force microscope (AFM) for
studies of the surface morphology of cells,
bacteria, viruses or even individual macromolecules e.g. DNA-protein complexes.
n A laser micro-dissection microscope
(Palm, Zeiss) for laser catapult capturing of
small tissue pieces, individual cells, microbes,
and chromosomes for subsequent PCR or
proteomics analysis.
n An IVIS biophotonic imaging instrument to trace luciferase labeled cells or
microbes in live experimental animals.
Laszlo Szekely and some of the members
of his group have developed new instruments
(programmable microscopes for automated
screening of 384 well plates), new image
analysis computer programs and a series of
automated microscopy based assays for highthroughput live cell imaging.
Using these assays it is possible to study
the effect of different drugs, antibodies, extracellular matrix components on the viability,
morphology and mobility of normal and
tumor cells as well as interaction with different microbial pathogens, immune effector
cells and/or stromal cells in multi-wavelength
time lapse assays.
Laszlo Szekely
Emilie Flaberg
Department of Microbiology, Tumor and Cell Biology – MTC
95
Core Facilities
MTC Research Facility
A cornerstone for research on infection and experimental cancer models
As a central Karolinska Institutet (KI) core
facility the MTC Research Facility has become
a cornerstone for research on infections and
experimental cancer models. In addition,
experimental mouse models for angiogenesis,
atherosclerosis, diabetes, rheumatism and
other diseases are of increasing importance
and are actively developed and explored. A
large collection of immunodeficient, gene
knockout, gene transgenic and other special
mouse strains are also available for research
purposes.
The facility is widely used by researchers
from MTC as well as researchers from other
KI departments. Several external researchers,
from other universities and from some private
companies, most of them linked to KI, use
the facility on a regular basis. In total, more
than 70 research groups perform their animal
experiments at the MTC Research Facility.
Strict routines
The facility has separate subunits for breeding, research, quarantine and biocontainment for experiments on infectious diseases.
Researchers have full access to the research
and biocontainment units after having
passed an introduction program at the KI
and locally.
Breedings of special mouse strains is
carried out in the breeding unit to which
only a limited number of animal care takers
have full access. Import and export of mice
is handled via the quarantine/reception unit.
Procedures for the daily handling of mice
follow the “3 Rs” principle of animal research:
“replacement, reduction and refinement”
in order to improve research quality and
increase animal welfare.
Before any animal experimentation can
start, all planned experiments have to pass
ethical clearance. In an application to the local
Ethical Committee, appointed by the government, a detailed scheme for the planned
procedures is declared and explained.
Researchers working at MTC have around
200 such individual ethical permits.
For import and export of mice special routines are followed. This includes evaluation
of health status reports and consulting KI’s
veterinarians. 1–2 times every year the animal
facility is inspected by representatives for the
county’s animal welfare unit.
Thanks to a strict entrance regulation
policy, training sessions and a strict import
policy for mice, the number of infections that
risk disturbing research have been kept very
low, especially in the breeding and research
centers.
Future development
A detailed plan for a major reconstruction of the MTC Research Facility has been
prepared to provide an even safer and more
efficient handling of animals. This includes
the construction of an air shower system for
access to the breeding and research units. In
addition, new washing machines for cages
and flasks will be installed and individually
ventilated cages (IVC) will be introduced in
all animal rooms.
New laboratories specially equipped for
studies on infections and cancer development
will be available. Hopefully the renovated
animal facility will be in operation during
2011–2012.
Anders Örn
Petter Höglund
The MTC Research Facility –
some features
n
A “core facility” for researchers at MTC,
KI and external users
n
High-quality laboratory technical assistance
n
A Cs-137 irradiation source for irradiation of cells and whole animals (transplantation studies)
n
Experimental infection models for virology, bacteriology and parasitology
n
Experimental cancer and angiogenesis models for studies of cancer
pathogenesis and new therapies
n
One of Sweden’s largest collection of
genetically modified ­
animals (> 100
strains)
n
Humanized mice for studying human
cells “in vivo” in a mouse model
n
Live-imaging to follow infections and/
or tumor growth in the l­iving mouse
(xenogenic-IVIS)
n
Visualization facility closely linked to
the animal house (KIVIF)
n
A special unit for research on germfree animals
Personnel
The Research
Facility’s IBL 637
irradiation unit.
96
Department of Microbiology, Tumor and Cell Biology – MTC
7Animal care takers
3Animal technicians
1Head of animal facility
1Compliance manager (Godkänd
föreståndare)
Core Facilities
The BSL-3 laboratory is a KI-core facility and an important platform to develop HIV research at the Karolinska Institutet.
HIV-laboratory
An efficient platform to perform HIV research
is a KI-core facility
and an important platform to develop HIV
research at the Karolinska Institutet (KI).
There are several groups operating within the
field of biology, virology and immunology of
HIV infection and these groups are located in
pre-clinical and clinical environments.
More than 150 PhD-theses have been
presented on the subject of HIV/AIDS from
KI during the last 20 years. Considering
the fact that there are so many established
research groups at KI and the Karolinska
University Hospital devoted to HIV work
this core facility will most likely be needed for
many years!
Due to safety aspects HIV work is
conducted at the BSL-3 laboratory level; it is
in fact desirable to have a laboratory devoted
exclusively to work with known HIV-contaminated material. The major hazard to laboratory personnel working with HIV infected
blood and body fluid is contamination of
hands and mucous membranes of the eyes,
nose and mouth. The MTC-BSL-3 facility is
a safe and efficient technical platform for HIV
research, for training and capacity building
within the frame of international collabo-
The BSL-3 laboratory
rations existing at the KI and Karolinska
University Hospital.
The users of this facility are already
involved in several international collaborations with countries heavily affected by HIV
infection and AIDS. These collaborations
are supported by the Swedish International
Development Cooperation Agency (Sida),
the World Health Organization, the European Commission and the Karolinska International Research and Training Committee
(KIRT). In addition to the academic partners,
also a few companies use the BSL-3 facility.
The users of the BSL-3 facility pay user fees
per hour. The fee is meant to cover material,
equipment service and the salary of a half
time technician.
The security rules established to work
within the BSL-3 laboratory ensures that the
HIV work is conducted safely. The approval
to work in the BSL-3 laboratory is periodically reviewed and authorized by the Swedish
Work Environment Authority.
Admittance to this facility is allowed only
following contact with responsible personnel, for teaching of related safety issues and
discussion and for practical introduction to
the facility. Before admission to the BSL-3
laboratory, all users need to attend a course
of three hours. To fresh up the competence,
each user attends a two hours course on the
simulation of accidents during work with
HIV each year. For convenient access, the
regulations and rules are summarized in the
booklet “Safety regulations at the BSL-3 safety
lab at MTC” and are the basis for the work in
the HIV laboratory to be followed by all users.
In general, the methods used in the BSL-3
facility aim at isolating and characterizing
HIV and studying immune responses to this
virus. Many of these analyses have relevance
to follow the course of HIV pathogenesis,
for the development of an HIV vaccine and
improved antiviral therapy.
The most common methods are: Isolation
of HIV from biological specimens; isolation
and culture of different cell subpopulations
from blood and biological specimens from
HIV infected patient; HIV Neutralization
assay to evaluate the titers of neutralizing
antibodies present in biological fluids of HIV
infected patients; HIV susceptibility and
resistance to different drugs.
Francesca Chiodi
Department of Microbiology, Tumor and Cell Biology – MTC
97
Interactions with the Outside World
During 2007–2009, MTC had a very active and
broad “out-reach” activity including appearance in TV and radio, in the general press, by
giving public lectures, by taking part in public
theatre and TV-productions and by the writing of popular science books.
”Research in Focus” (Forskning i Fokus)
is an evening activity for the general public
interested in our research areas and their
implications for society and its citizens. It is
announced in daily newspapers and on websites. Usually the audience fills a large lecture
hall. In the autumn 2007 we had an unusual
experience. One of our topics on “Gut feeling – the gut and its microbes” raised a crowd.
As a result some 100 persons could not be
accommodated and we repeated the event
one month later – with the same results! So
we had a third evening with the same theme in
early 2008 in the largest lecture hall of Karolinska Institutet (KI). These three occasions
attracted a crowd of close to 800 persons – a
new record for this activity.
Another type of activity is the “What is
life”-lectures organized together with the
Books written by MTCers
Magen
Peter Benno,
Ingemar Ernberg,
Claude Marcus,
Tore Midtvedt,
Roland ­Möllby,
Elisabeth Norin,
Torgny Svenberg
Meteorer
Georg Klein
Framtidens ­farliga
smitta
Britta ­Wahren,
Patrik Wahren
98
Department of Microbiology, Tumor and Cell Biology – MTC
KI Cultural Board. This series is
open to the general public and has
become an important arena for
going beyond our everyday scope in
science.
During 2007–2009 we had 27
invited speakers including Craig
Venter, among the first to sequence
the human genome; Peter Gärdenfors, professor of cognitive science
and one of Scandinavia’s most
notable living philosophers; Svante
Pääbo, director at the Max Planck
Institute for Evolutionary Anthropology; James Watson, codiscoverer of the DNA structure; Matthias
Uhlén, leader of the Human Protein
Resource Project; Göran Burenhult,
a Swedish archeologist and Edward
O Wilson, a socioecologist.
From Cirkus Cirkör’s “Inside out”. Photo Mattias Edwall.
MTC faculty members also regularly teach natural science classes at
the high school level in two schools
normal flora of the gut (the “2 kg-gruppen”).
outside Stockholm. For some students this
In addition, Georg Klein has written a book
includes project work with MTC scientists. In
about brilliant, eccentric scientists.
addition, during every year the students have
A number of public appearances have
a thematic week together with the KI faculty.
been organized at the public libraries of
First year was “Medicine through history”,
Stockholm. Most noticeable were the three
second year “World health” and third year
discussion evenings at the Stockholm City
“Cancer”.
library during spring of 2009 organized
together with the Karolinska University
Press AB under the thematic name “Research
Outstanding cancer scientist
important for life”.
In connection with the George and Eva Klein
Award 2008 a public lecture was given by the
outstanding cancer scientist Robert A. WeinCirkus Cirkör
berg, Whitehead Institute, Cambridge (US)
MTC has been involved in the Cirque Noveau
on “How do we deal with the cancer epidemgroup “Cirkus Cirkör” – to provide ideas and
ics”. Another public lecture was given by the
knowledge in modern biology, which contribP.R. China Minister of Health, former cancer
uted to their two shows “99% unknown” 2007
scientist Professor Chen Zhu on the adoption
and “Inside-Out” 2009, performed at the
of traditional Chinese medicine to modern
“House of dance” in Stockholm but also tourmedicine and the coming Chinese health care
ing world-wide.
reforms which he is launching.
During 2008 and 2009 several of our
MTC faculty participates actively in the
faculty members and students were involved
Karolinska University Press AB popular book
in the Lennart Nilsson-Mikke Agaton producproduction, including co-authorships in the
tion “A journey to the core of life”, a 2 hour TV
following books (titles translated): Prostate
production which was shown in 2010. IngeCancer, Breast Cancer, Dangerous infecmar Ernberg was the scientific advisor of this
tions of the future, The gut and its bacteria
project.
(”Magen- bakterier, buller och brak”). The
Ingemar Ernberg, chairman
latter was initiated from MTC projects on the
of the KI Cultural Board
Professor Robert A Weinberg and HRH Crown Princess Victoria together with Eva
and Georg Klein.
Photo: Ulf Sirbjorn
Professor Robert A Weinberg received the Visiting Professor Award
from the hands of HRH Crown Princess Victoria. Photo: Ulf Sirbjorn
Georg and Eva Klein Foundation
In support of innovative research
The “Georg and Eva Klein Foundation”
The board of the foundation has eight
members representing both science and society. The board convenes 4–5 times per year
and had the following composition during
2009:
(GEKS) was established in 2005 initiated by
researchers from MTC in order to celebrate
the 80th birthdays of Georg and Eva Klein, the
founders of the Department of Tumor Biology at the Karolinska Institutet in 1960.
Ebba Lindsö, Entrepreneur
Georg and Eva’s ground-breaking reCarl Fredrik Sammeli – Founder, Prime
search, from the ’60s until today, contributed
Group
a number of very important scientific findAlf Svensson – Member of the EU parliament
ings in the areas of tumor immunology and
Magnus Uggla – Executive Vice President,
tumor biology. To mention a few, the capacHandelsbanken
ity of the immune system to recognize and
Marie Arsenian Henriksson – Head of MTC
react against certain tumors, the role of EBV
Ingemar Ernberg – Professor
in lymphoid malignancies, the importance
Klas Kärre – Professor
of chromosomal translocations for tumor
Anders Örn – Professor (Chairman)
development, identification of tumor suppressor genes and important aspects of EBVhost interactions for viral latency and tumor
Those who have and have had the privilege to
development.
be trained and to mature scientifically under
The main objective of the foundation
the auspices of Georg and Eva Klein often talk
is “to support biomediabout the unique culture created
cal research in the spirit
in their laboratories. To capture
of Georg and Eva Klein.”
the “Klein spirit” in words is not
Funds are collected in oreasy. One important factor is the
der to support research
actual mentoring process that
on cancer and cell biology,
is performed in a very personal
immunology, genetics and
manner, where curiosity is an
microbiology. The focus is
important driving force.
www.gekleinfoundation.org
on:
Another success factor is to let
the most junior students explain
n Basic research with the potential to deand discuss their research projects directly
with very experienced visiting scientists. This
velop knowledge that can lead to medical
exposure fosters confidence and triggers a
applications
n Knowledge sharing through international
curiosity-driven attitude to science.
The spirit of learning by doing is also
scientific exchange and networking
n Science with a humanistic perspective
important where trial and error(s) are neces-
sary components. The “Klein spirit” also tries
to support researchers whose projects contain
more than data collection and potentially can
lead to synergy between disparate fields of
science.
World famous research
In 2008 two scientific prizes were awarded
by the foundation. The “Visiting Professor Award” was given to Robert A Weinberg
working at MIT, Cambridge, USA. His world
famous research is focused on the genetic
basis of cancer development and metastasis.
In addition to his research achievements
Robert Weinberg has written a popular textbook with the title “The Biology of Cancer”.
During a hectic week in Sweden he gave a most
appreciated series of lectures for researchers and students. In addition, he gave an
outstanding presentation on “The future of
cancer – research and treatment” for a layman
audience. Dr Weinberg also held several highly
interactive seminars with junior scientists on
various themes in cancer biology.
Dr Theresa Vincent received the “Junior
Scientist Award” at the same occasion for her
excellent research on the effect of cell signaling on tumor cell behavior. This stipend
partly supported her research at the Cornell
University in New York.
The actual prize ceremony was held at
the Nobel Forum, KI in the presence of HRH
Crown Princess Victoria as well as many
researchers and some of the generous donors
to the foundation.
Anders Örn
Department of Microbiology, Tumor and Cell Biology – MTC
99
MTC Organization
MTC has a flat organization with around 35
research groups, each led by a group leader,
a professor or a senior researcher. The group
leader is responsible not only for the scientific
work but also for the economy and the staff in
the group.
Marie Arsenian Henriksson has been the
Head of the Department since October 2006.
She has since then been appointed by the KI
President for a second three-year period.
The Head of Department has assigned a
management group consisting of the two vice
chairmen, the head of administration (AC)
and four group leaders. Professor Staffan
Normark and Professor Klas Kärre are vice
chairmen and Åsa Edström has been the head
of Administration since August 2008.
Joint
Influence
Group
Equality
and Diversity
Group
Department
Council
Future
Faculty
Management Group during 2009
Work
Environment
Group
Marie Arsenian Henriksson (Chair)
Åsa Edström
Annelie Brauner
Klas Kärre (Vice Chair)
Staffan Normark (Vice Chair)
Francesca Chiodi
Gunilla Karlsson-Hedestam
Pontus Aspenström
Faculty
Advisory bodies
Importantly, there are a number of advisory
bodies, including the Department Council, the Faculty (the group leaders), the Post
Graduate Research Committee, the Work
Environment Group, the Equality and Diversity Group and the Undergraduate Council.
The Department also has a Joint Influence
Group, a Future Faculty, and a Student Council. In addition, there is an external Scientific
Advisory Board (see box), which interacts
with the management of MTC to discuss
issues of importance for the long-term development of the department.
Support for the research groups
The main focus of the Administration and
The Service Unit is to support the research
groups when it comes to economy, staff
administration and different service functions.
In order to streamline the service and
infrastructure at MTC previous units for
service, IT and shops for laboratory material were merged into one, the Service Unit,
during 2009 . A head for this new unit was
subsequently recruited, Petra Hartley.
The department also has several core
facilities Including KIVIF, the FACS facility,
the Research Facility and the HIV laboratory
supporting the ongoing science.
Åsa Edström
100
Department of Microbiology, Tumor and Cell Biology – MTC
Third Level
(Postgraduate)
Research
Committee
Student
Council
Second Level
(Undergraduate)
Research
Committee
Scientific Advisory Board
Xin Lu, professor and director, Ludwig Institute for Cancer
Research, Oxford, Great Britain.
Peter H. Krammer, professor and Head of the Tumor
­Immunology Program, German Cancer Research Center
(DKFZ), Heidelberg, Germany.
Olle Stendahl, professor, Linköping University, Linköping,
Sweden.
MTC Staff
Technical staff
Professor Xin Lu.
Female
Male
Total
39
11
50
Administrative staff
18
1
19
Postgraduate students
90
55
145
Postdocs
20
13
33
4
6
10
11
9
20
Research assistants
Researchers
Senior lecturers
1
1
2
Professors
4
16
20
187
112
299
TOTAL
Economy
Result (in kSEK)
Income of governmental grants
Income of fees
Income of external grants
Internal incomes
2009
2008
2007
2006
63 735
56 735
55 329
58 613
4 786
4 989
5 399
6 737
110 714
102 258
101 330
103 804
12 986
8 481
21 113
18 900
SUM OF INCOMES
192 221
172 463
183 171
188 054
Personnel costs
-111 246
-101 151
-99 537
-102 802
Premises costs
-32 434
-31 178
-30 964
-31 509
Operational costs
-47 600
-38 786
-38 200
-34 237
-3 714
-3 215
-3 639
-3 867
-194 994
-174 330
-172 340
-172 415
Financial incomes
216
1 895
1 179
396
Financial costs
-79
-59
-49
-37
FINANCIAL NET
137
1 836
1 130
359
Change of capital
-2 636
-31
11 961
15 998
Depreciation
SUM OF COSTS
Development of MTC incomes
Income according to contributor in 2009
kSEK 200
Foundations
and organizations
150
Internal income
Foreign research
grants
Other
govermental
contributions
100
KI govermental
grants
50
National
research
councils
0
94 95 96 97 98 99 00 01 02 03 04 05 06 07 08 09
Sum of incomes
Internal incomes
Income of contributions
Income of fees
Income of governmental
grants
Department of Microbiology, Tumor and Cell Biology – MTC
101
Contributors and List of Financiers
We would like to thank our financiers for their valuable support to our research!
The eight major external contributors 2007
The eight major external contributors 2009
ksek
21 649
Swedish Research Council (VR)
40 390
European Union (EU)
15 214
Swedish Cancer Society (CF)
19 613
Swedish Cancer Society (CF)
15 144
European Union (EU)
16 872
10 531
Swedish Foundation for Strategic Research (SSF)
11 500
Swedish Institute for Infectious Disease Control (SMI)
Swedish Institute for Infectious Disease Control (SMI)
10 580
The Swedish Agency for Innovation Systems (VINNOVA)
4 207
3 500
The Swedish International Development Cooperation Agency (Sida)
3 981
The Swedish Childhood Cancer Foundation (BCF)
3 397
The Swedish Childhood Cancer Foundation (BCF)
3 226
Torsten and Ragnar Söderberg Foundation (TRSS)
2 432
Torsten and Ragnar Söderberg Foundation (TRSS)
3 000
The Swedish International Development Cooperation Agency (Sida)
–
–
–
–
–
–
–
–
–
–
–
–
–
Accuro AB
Actar AB
Active Biotech AB
AFA Insurance
Affibody Biotechnology AB
Åke Wiberg Foundation
Alex and Eva Wallström Foundation
Animal Protection Agency
Aprea AB
Arla AB
AstraZeneca AB
Athera Biotechnologies AB
Aventis Pasteur SA
–
–
–
–
–
Bill & Melinda Gates Foundation
Binex Company
BioInvent International AB
Biostapro AB
Biovitrum AB
– Cancer and Allergy Fund
– Cancer Association in Stockholm
– Cancer Research Funds of
Radiumhemmet
– Cancer Research Institute
– Capio Research Foundation
– Cardoz AB
– Carl Trygger Foundation
– Cederberg Foundation
– Clas Groschinsky Memory
Foundation
– Clinical Gene Networks AB
– DAKO AB
– Diamyd Therapeutics AB
– Dilafor AB
– Ekhaga Foundation
– Euro-Dignostica AB
– European Union (EU)
–
–
–
–
–
102
ksek
Swedish Research Council (VR)
German Cystic Fibrosis Foundation
Glaxo AB
Golje Memorial Fund
Göran Gustavsson Foundation
Harvard University
– Henning and Ida Persson Research
Foundation
– High Q Foundation
– Human Frontier Science Program
–
–
–
–
–
–
IMED AB
Immune Tolerance Network
Index Pharmaceutical AB
Independent Pharmaceutica AB
Ingabritt och Arne Lundberg
Research Foundation
Innogene-Kalbiotech Company
INTAS International Association
International AIDS Vaccine
Initiative
IPULS AB
–
–
–
–
Jeansson Foundation
Jerring Foundation
Jochnick Foundation
Julin Foundation
–
–
–
– Karo Bio AB
– Karolinska Institutet Foundations
and Funds
– Knut and Alice Wallenberg
Foundation
– King Gustaf V Jubilee Foundation
– Knowledge Foundation (KK)
– Knut and Alice Wallenberg
Foundation
– Konsul TH C Berghs Foundation
–
–
–
–
–
Länsförsäkringar Alliance
Lars Hierta Memory Foundation
LipoPeptide AB
LTB4 Sweden AB
Ludwig Institute for Cancer
Research
– Lundberg Research Foundation
–
–
–
–
–
–
Mabtech AB
Magn. Bergwalls Foundation
Mälardalen University
Mary Beves Foundation
Maths O Sundqvist Foundation
Max-Planck Institute
Department of Microbiology, Tumor and Cell Biology – MTC
– Nancy Lurie Marks Family
Foundation
– Nanna Svartz Fund
– National Association for Kidney
Diseases
– National Board of Health and
Welfare Foundation
– Norwegian Institute of Public Health
– National Institutes of Health (NIH)
– Nobel Foundation
– NOLabs AB
– NordVacc AB
– Novo Nordisk AB
– O.E. & Edla Johansson Veterinary
Foundation
– Olle Engkvist Foundation
– Oncoreg AB
– Petrus and Augusta Hedlund
Foundation
– PhPlate Microplate Techniques AB
– Physicians against AIDS Research
Fund
– Q-Med AB
– Robert Lundberg Memorial Fund
– Royal Institute of Technology (KTH)
– Royal Swedish Academy of
Sciences (KVA)
–
–
–
–
–
–
–
–
–
–
S*Bio Ltd
Scandinavian Clinical Nutrition AB
Scripp Research Institute
Sigurd and Elsa Goljes Foundation
Swedish Agency for Innovation
Systems (VINNOVA)
Swedish Board of Agriculture
Swedish Cancer Society (CF)
Swedish Childhood Cancer
Foundation (BCF)
Swedish Diabetes Association
Swedish Foundation for
International Cooperation in
Research and Higher Education
(STINT)
– Swedish Foundation for Strategic
Research (SSF)
– Swedish Heart-Lung Foundation
– Swedish Institute (SI)
– Swedish Institute for Infectious
Disease Control (SMI)
– Swedish International Development
Agency (Sida)
– Swedish Medical Association (SLS)
– Swedish National Board of Forensic
Medicin
– Swedish Nutritional Research
Foundation
– Swedish Orphan AB
– Swedish Research Council (VR)
– Swedish Research Council
FORMAS
– Swedish Rheumatism Association
– Swedish Society for Medical
Research (SSMF)
– Swedish Society of Medicine
– Swedish University of Agricultural
Sciences
– Thymed Company
– Tobias Foundation
– Torsten och Ragnar Söderberg
Foundation (TRSS)
–
–
–
–
–
Unimed Innovation Company
United Way International
University of Bergen
University of California
University of Rome, Tor Vergata
– Vaccine Research International Plc
– Virtual Genetics AB
– Viscogel AB
– WennerGren Foundation
– World Health Organization (WHO)
Location of MTC
Picture: John Sennett
Mailing Address
Visiting Address
Phone
Website
MTC, Karolinska Institutet
FE 280
SE-171 77 Stockholm
Sweden
Nobels väg 16 or Theorells väg 1
KI Campus, Solna
+46 (8) 524 8 66 78
http://ki.se/mtc
Clinical Microbiology
Laboratory building L2:02
Karolinska University Hospital, Solna
SE-171 76 Stockholm
Sweden
Karolinska University Hospital, Solna
Sjukhusringen 13
Laboratory building L2:02
+46 (8) 517 735 88
Visitors to MTC
From Arlanda Airport take the airport coach to Sankt Eriks­plan and bus 3, 77 or 507 to
bus stop “Karolinska Institutet” (4th stop). Walk across the campus to Nobels väg.
From the Central Train Station take bus 47 to Tomteboda
(end stop). Walk across the campus via Berzelius väg and
Nanna Svartz väg to Nobels väg.
By car from E4 North: follow the signs to Karolinska University Hospital (Karolinska
sjukhuset). Take exit 165 and turn immediately left. Follow Karolinska vägen until it
becomes Tomtebodavägen. Take then the first street to the left and find Nobels väg 16 on
the left after 50 meters.
From the city: follow Torsgatan and Solnavägen and turn left to Tomte­bodavägen.
By car from E4 South: Take the right lane towards S:t
Eriks­plan. Continue onto Norrtull. Take the right lane at the
Norrtull exit and turn right onto the bridge to Solna. Continue for about 600 metres and turn left into Tomtebodavägen
and then left into Nobels väg.
Department of Microbiology, Tumor and Cell Biology – MTC
103
Fungal cellulase binds to Acanthamoeba cell wall. Artistic interpretation by Ewert Linder adapted from: Appl Environ Microbiol. 75, 6827-30, 2009.