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Department of Neurology · THE Neuroscience Centre ·
Copenhagen University Hospital · Rigshospitalet · Copenhagen, Denmark ·
www.ms-research.dk
Danish
Multiple Sclerosis Center
Annual Report
2009
DANISH Multiple sclero sis CE N TER ann ual report 2009
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DMSC staff seminar 2009.
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DANISH Multiple sclerosis CEN TER annual report 2009
Table of contents
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A short review of 2009 in the Danish Multiple Sclerosis Center (DMSC)
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DMSC missions and aims
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About the DMSC
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Organization
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Research activities 2009
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Clinical research
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Neuroimaging
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Neurogenetics
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Neuroimmunology
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Neuropathology
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Routine analyses in Neuroimmunology Laboratory
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Scientific publications 2008-2009
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Prizes
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Honorary offices
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Scientific collaboration
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Acknowledgements
Annual
Report
2009
DANISH Multiple sclero sis CE N TER ann ual report 2009
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Publisher:
Danish Multiple Sclerosis Center
Authors:
Per Soelberg Sørensen
Morten Blinkenberg
Finn Sellebjerg
Annette Oturai
Helle Bach Søndergaard
Photos:
Peter Fischer
Concept, design, graphic
­production and print:
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Njalsgade 19 D
DK-2300 København S
Tlf.: +45 89 39 88 33
www.stibozone.com
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DANISH Multiple sclerosis CEN TER annual report 2009
Professor Per Soelberg Sørensen,
MD, DMSc
Director of the Danish Multiple
Sclerosis Center (DMSC)
A short review of 2009
in the Danish Multiple
Sclerosis Center
In 2009 the number of patients in the Danish Multiple Sclerosis Center (DMSC) increased to 1777. To meet the challenge of
providing care of an increasing number of MS patients referred
for diagnosis and therapy, in particular for treatment with
­Tysabri infusions, we employed an additional staff neurologist,
MS nurse and secretary to the MS Clinic. By the end of 2009
approximately 300 patients were receiving monthly infusions
of Tysabri and it has been a challenge to organize the treatments due to the limited space in the MS Clinic facilities.
In 2009 we completed two multi-center, international trials
organized and directed from DMSC. These trials showed that it
is possible to achieve significant improvement in the efficacy of
interferon-beta by giving monthly pulses of methylprednisolone as add-on therapy.
We have expanded our international research collaboration
in clinical research, neurogenetics, neuroimmunology and MS
pathology. We have published a series of scientific papers on
response of the immune system to various treatments. These
papers have extended our understanding of the deleterious effects of antibodies to interferon-beta and a paper has reported
the first series of patients treated with Tysabri outside a clinical
trial setting. We have published papers together with Austrian
collaborators showing that compartmentalized central nervous
system inflammation is closely related to neurodegeneration
giving us the hope that therapies able to eliminate the inflammation in the central nervous system will stop the degenerative
processes in the brain and spinal cord.
I hope that you will enjoy reading the annual report of DMSC.
Per Soelberg Sørensen
DANISH Multiple sclero sis CE N TER ann ual report 2009
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DMSC missions and aims
The mission of Rigs­hospitalet is to be the
leading hospital in Denmark for patients in need
of highly specialized treatment
The missions of the
­Danish Multiple Sclerosis Center (DMSC) are:
To be the leading multiple sclerosis (MS) center
in Denmark
To be at the forefront of highly specialized
management of MS
To carry out research and development in MS
at an advanced international level
To collaborate scientifically and exchange
knowledge in MS research
To educate staff to a highly specialized level in
their relevant fields
To contribute with professional advice on MS
to the healthcare community
To meet people with MS at their terms with
openness and respect
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DANISH Multiple sclerosis CEN TER annual report 2009
The aims of
the DMSC are:
To provide the optimal interdisciplinary patient
care to all MS patients in the region and to
patients from other regions in need of highly
specialized therapy
To carryout high quality research in MS with
focus on clinical research, new therapies, MS
genetics, neuroimmunology and MS pathology
To teach undergraduate students and PhDstudents and stimulate their interest in MS
research
To educate post docs, MS physicians, nurses,
secretaries and other professionals to a high
level of knowledge of MS in their relevant
expert fields
To lead the national research in Denmark in
partnership with other Danish researchers and
to establish a broad international collaboration
with MS research groups in Europe and from
overseas.
DANISH Multiple sclero sis CE N TER ann ual report 2009
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About
The Danish Multiple
Sclerosis Center
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DANISH Multiple sclerosis CEN TER annual report 2009
DANISH Multiple sclero sis CE N TER ann ual report 2009
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The Danish Multiple Sclerosis Center
About The Danish Multiple
Sclerosis Center
The Danish Multiple Sclerosis Center (DMSC) is
composed of the MS Clinic and the MS Research
Unit.
The MS Clinic is located on the 8th floor of the
main complex of Rigshospitalet. The MS Clinic
comprises offices of the professor and consultants,
the reception desk and secretary office, the nurses’ offices, the consultation rooms as well as the
facilities for intravenous therapy with Tysabri and
rooms for invasive procedures. The MS Clinic provides a multi-disciplinary care for almost 1800 MS
patients of whom the majority receive diseasemodifying therapy dispensed by the MS Clinic.
Apart from disease modifying therapy patients are
offered symptomatic therapy for spasticity, bladder and bowel disturbances, pain and cognitive
symptoms. The medical staff comprises a professor, 4 consultants and 3 staff neurologists; there
is one leading nurse and 6 MS specialist nurses, 3
secretaries, a neuropsychologist, a physiotherapist
and a medical social counselor.
Approximately half of the patients in the MS
Clinic are from our local Copenhagen region and
the other half are patients referred for highly specialized therapy from the neighbouring regions all
over Zealand. The MS Clinic has been appointed
by the National Board of Health to perform highly
specialized therapy such as strong immunosuppression and Tysabri therapy. In addition patients
are referred for experimental therapy with highly
specific monoclonal antibodies. Further, the MS
Clinic provides highly specialized treatment of
spasticity with an intrathecal baclofen pump. In
collaboration with pediatricians children and adolescents with MS from Eastern Denmark are also
treated at the MS Clinic.
It is the aim of the MS Clinic to provide high
quality multi-disciplinary care for all our patients
with openness and respect.
The MS Research Unit is partly located in
the proximity of the MS Clinic and partly in the
Michaelsen Building 63. Clinical research has to
be performed in close proximity to the MS Clinic
and offices for 3 research nurses and 1 research
secretary are embedded in the MS Clinic. The
remaining part of the MS Research Unit with the
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DANISH Multiple sclerosis CEN TER annual report 2009
Neuroimmunology Laboratory is located on the
first floor in the Michaelsen building 63 and in the
basement of building of 93. The facilities contain
offices for the research staff and laboratories. The
laboratories contain an 8-color flow cytometer,
facilities for doing real-time polymerase chain­
reaction (PCR), a neurogenetics laboratory for
DNA preparation and facilities for making routine
laboratory tests. The focus of the research is clinical research including neuroimaging, neuroimmunology, neurogenetics and neuropathology of MS.
ACtive patientS iN DMSC
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Organization
Director: Professor Per Soelberg Sørensen
MS clinic
MS Research Unit
Neurologists
Professor Per Soelberg Sørensen
Consultant Morten Blinkenberg
Consultant Finn Sellebjerg
Consultant Annette Oturai
Consultant Karen Schreiber
Staff neurologist Ana Voldsgaard
Staff neurologist Lars Pindborg
Staff neurologist Melinda Magyari
Staff neurologist Henrik Mathiesen
Clinical research
Professor
Per Soelberg Sørensen
Consultant Karen Schreiber
Staff neurologist Ana Voldsgaard
Staff neurologist
Melinda Magyari
Neuropsycologist
Agnete Jønsson
PhD student Stephan Bramow
PhD student Lars Börnsen
PhD student Jeppe Romme Christensen
PhD student Dan Hesse
PhD student Rikke Ratzer
Junior physician Morten Møller
MS nurses
Leading nurse Anne Hansen
Dorte Stauning Rasmussen
Anette Husted Pedersen
Lene Almind
Julie Yoon S. Moberg
Louise Nathalie Christiansen
Maj Daae Christensen
Sidsel Nielsen
Secretaries
Annette Larsen
Malene Møllesøe
Maria Brændbjerg
Helle Vilhhelmsen
Research nurses
Vibeke Jespersen
Joan Pietraszek
Sidsel Nielsen
Research secretary
Annette Larsen
Neuroimaging research
Consultant
Morten Blinkenberg
Staff neurologist
Henrik Mathiesen
Genetic research
Consultant Annette Oturai
Senior research fellow
Helle Bach Søndergaard
Junior research fellow
Morten Møller
Neuropsychologist
Agnete Jønsson
Physiotherapist
Lis Albrechtsen
Medical social counselor
Keld Nissen
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DANISH Multiple sclerosis CEN TER annual report 2009
Neuroimmunology
research
Ass. professor Finn Sellebjerg
Senior research fellow
Helle Bach Søndergaard
PhD students
Dan Hesse
Lars Börnsen
Jeppe Romme Christensen
Rikke Ratzer
Junior research fellow
Marianne Hansen
Neuropathology
research
Professor
Per Soelberg Sørensen
Ph.d. student
Stephan Bramow
Neuroimmunology
Laboratory
Laborarory leader
Poul Erik Hyldgaard Jensen
Laboratory technicians
Leading Laboratory technician
Joy Mendel-Hartvig
Marie Koefoed
Anne Mette Hedegaard Nielsen
Michael Jensen
Vibeke Fuglholt
Professor
Per Soelberg
Sørensen
Ass. professor
Finn Sellebjerg
Consultant
Morten
­Blinkenberg
Consultant
Annette Oturai
Consultant
Karen Schreiber
Staff neurologist
Ana Voldsgaard
Staff neurologist
Henrik
Mathiesen
Staff neurologist
Melinda
Magyari
Laboratory
leader Poul Erik
Hyldgaard
Jensen
Senior research
fellow
Helle Bach
Søndergaard
Leading laboratory technician
Joy MendelHartvig
Leading nurse
Anne Hansen
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Research
activities 2009
Clinical research
Clinical research
Neuroimaging
Neurogenetics
Neuroimmunology
Neuropathology
Routine analyses in Neuroimmunology Laboratory
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DANISH Multiple sclerosis CEN TER annual report 2009
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Research activities 2009
Clinical research
Clinical research group:
Per Soelberg Sørensen, Morten Blinkenberg,
Finn Sellebjerg, Annette Oturai, Ana Voldsgaard,
Karen Schreiber, Henrik Mathiesen,
Melinda Magyari, Dan Hesse, Stephan Bramow,
Lars Börnsen, Jeppe Romme Christensen,
Agnete Jønsson, Vibeke Jespersen, Joan Pietraszek,
Sidsel Walther Nielsen, Anne Hansen,
Annette Larsen
Therapeutic trials
The results of two international multi-center
clinical trials managed by DMSC using monthly
pulse therapy of methylprednisolone tablets as
add-on therapy to interferon-beta were presented
at major MS congresses in 2009. The NORMIMS
trial showed that in patients with breakthrough
disease on interferon-beta 1a treatment add-on
therapy with methylprednisolone tablets 5 days a
month reduced the monthly relapse rate by 62%
compared to add-on of placebo and reduced the
accumulation of lesions on MRI. These results were
corroborated in the MECOMBIN study of previously untreated patients showing that the combination of interferon-beta 1a and methylprednisolone
tablets 3 days monthly reduced the relapse rate by
38% and showed beneficial effect in MRI outcomes
compared to interferon-beta and placebo.
Two other large combination trials are currently
directed from DMSC. The SIMCOMBIN study is a
Nordic multi-center study of Simvastatin as addon therapy to interferon-beta in de novo treated
The MS Clinic is
located on the
8th floor of the
main complex
of Rigshospitalet.
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DANISH Multiple sclerosis CEN TER annual report 2009
patients with relapsing-remitting MS. More than
300 patients have been recruited and the trial will
be completed by the end of April 2010. The RECYCLINE study is a European multi-center study exploring the effect of Minocycline as add-on therapy
to interferon-beta in de novo treated patients.
Recruitment of 320 patients has been completed
and the results will be available in 2012.
We are currently performing three single–center
studies: A study of erytropoietin (EPO) in patients
with progressive MS; a small safety study of treatment with eggs of the pig whipworm (Trichuris
suis); and we have as the first center initiated an
exploratory trial of natalizumab (Tysabri) in patients with progressive MS.
Neutralizing antibodies (NAb) against
biological therapies
In a study using affymetrix gene chips we showed
that interferon-beta treated NAb-positive patients,
in whom MxA could not be induced by interferonbeta, had no residual bioactivity present and were
virtually untreated.
DMSC collaborates with an a EU supported
European study of neutralizing antibodies against
interferon-beta (NABINMS study). We are currently analyzing data from population-based cohorts of
patients from Denmark, Spain, The Netherlands and
The Czech Republic. Further, the NABINMS study
has introduced a new assay for neutralizing antibodies based on a Luciferase reporter gene under
control of interferon-beta, and this assay has been
adapted for routine measurements of neutralizing
antibodies in DMSC.
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Research activities 2009
Neuroimaging
Neuroimaging research group:
Morten Blinkenberg, Henrik Mathiesen,
Per Soelberg Sørensen
PET
During the last decades, functional imaging have
improved our understanding of the neurodegenerative processes in MS.
Former positron emission tomography (PET)
studies, carried out in the Danish Multiple Sclerosis
Center, have shown that cerebral activation in MS
patients, is severely reduced as a consequence of
disease progression.
Our current PET studies focuses on the early relapsing remitting phase of the disease and the corresponding changes in cerebral activation. Preliminary
results show, that reductions in PET is associated
with neuronal dysfunction or loss, measured by
magnetic resonance spectroscopy (MRS). In this way
there seem to be a close relationship between these
two measures of cerebral neuronal integrity in MS.
MRI
In collaboration with Danish Research Centre
for Magnetic Resonance (DRCMR), Hvidovre
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Hospital we have previously shown, that
measurements of MRS correlate with cognitive
dysfunction in MS. This cohort has now been reevaluated and analyzed for longitudinal changes
in MRS. Preliminary data show, that MRS decrease in the early stage of MS, predicts disease
progression after 5 years. MRS may therefore
be used as a clinical tool, to determine disease
course in MS.
Another study focuses on the pathophysiological mechanisms underlying changes in cerebral
activation in MS patients. Resting-state fMRI is a
new approach to assess functional brain connectivity by measuring the synchronous fluctuation
in the blood-oxygen-level dependent signal between remote brain regions at rest. DRCMR uses
this method to study, whether changes in functional connectivity of the motor network, reflect
disease severity in patients with MS. Brain
functional connectivity is investigated in a crosssectional study of 42 patients and furthermore in
a longitudinal study of 12 patients evaluating the
temporal dynamics during a relapse and clinical
remission. Patient recruitment has been completed during year 2009 and elaborated statistical
analysis is ongoing.
Neurogenetics
Neurogenetics research group:
Annette Bang Oturai, Helle Bach Søndergaard,
Finn Sellebjerg, Julie Maria Hejgaard
The ultimate cause of multiple sclerosis (MS) is still
unknown. The probable cause is thought to be a
combination of hereditary factors, an environmental trigger and a defect in the immune system. From
family studies we have known for many years that
genes plays a role.
Identifying the involved genes is a complex challenge, because no single Mendelian locus causes
MS. Through decades scientists have worked to
find gene variations tied to MS. Genome linkage
screens have failed finding major genes except
genes of the HLA-complex, where linkage to DR2
(HLADRB1*15) is strongest in Northern Europeans. The HLA-DR2 association to MS was already
identified back in 1973. A break-through took
place in the summer 2007, when three new studies
showed that the IL7 receptor and IL2receptor genes
were important risk genes of MS. Both genes are
important for the T cell regulation, and thus the
immune system. Since then, several new genes
have been confirmed. Presently, the Wellcome Trust
Case Control Consortium is investigating the largest
set of MS cases and controls (11.000/11.000) by a
500.000 SNPs chips. Results are expected later this
year. We are taking part in this large international
genetic collaboration through our membership of
the IMSGC (International Multiple Sclerosis Genetic
Consortium).
For more than 15 years the MS Genetic Group
at Rigshospitalet have collected DNA, and today
we have DNA from more than 1800 Danish MS
patients and 1200 controls, all kept in the “Danish
Multiple Sclerosis Biobank” located at our department at Rigshospitalet. In order to increase the
sample size for genetic testing, we have participated in the “Nordic MS Genetic Network” since 1994,
and today the Nordic material consists of 5000 MS
cases and 5000 controls. Local research is focused
on the candidate gene approaches and the genetic
influence on the differences in treatment response.
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Research activities 2009
Neuroimmunology
Neuroimmunology research group:
Finn Sellebjerg, Helle Bach Søndergaard,
Poul Erik Hyldgaard Jensen, Dan Hesse,
Jeppe Romme Christensen, Lars Börnsen,
Rikke Ratzer, Per Soelberg Sørensen
Immunological studies
The interplay between the different types of
immune cells in relapsing-remitting MS is only
incompletely understood. Furthermore, the role
of immune cells in the pathogenesis of the slowly
developing deterioration in chronic progressive MS
is a matter of debate. Some researchers consider
immune activation to be of crucial importance in
the pathogenesis of progressive MS, others consider
progressive MS to result from slowly developing,
neurodegenerative processes that are initiated by
previous, inflammatory insults rather than by active
inflammation.
We are using a combination of immunological
studies, addressing the reactivity of subsets of T
cells to autoantigens expressed in the brain and spinal cord, studies addressing the role of other leukocytes in the activation of T cells, and studies of the
effect of immunomodulatory treatment to further
our understanding of the role of immune activation in different disease stages in MS. Initial studies
using these techniques, conducted in collaboration
with the Institute for Inflammation Research at the
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DANISH Multiple sclerosis CEN TER annual report 2009
Copenhagen University Hospital Rigshospitalet,
have provided evidence that active MS is associated
with the activation of a recently discovered subtype
of T cells termed Th17 cells. We are now studying
the nature of these cells in more detail.
In other studies we use flow cytometry, which is
a method that allows the analysis of the phenotype
of individual, circulating lymphocytes by measuring
the binding of fluorochrome-coupled, monoclonal
antibodies to distinct molecules. This technique is
well suited for studying the activation status of
subsets of lymphocytes. By combining the analysis
of different molecules expressed on the cell surface,
these cells can be divided into a large amount of
functionally relevant subtypes, which can be quantitated both in relative and absolute terms. We have
previously reported that flow cytometry can detect
changes in the activation of circulating T cells and
monocytes that differ in relapsing-remitting and
secondary progressive MS. We are now exploring
the extent to which such changes can be observed
in other lymphocyte subsets, and whether they are
associated with changes in the expression of cytokines and intrathecal immune activation that may
reflect disease activity in progressive MS.
Molecular biology
The functional activation of immune cells requires
complex interactions between different immune
cells and coordinated, tightly regulated expression of numerous different molecules within single
cells. DNA arrays provide a platform for unbiased
studies of gene expression in different disease
states and for studying the effect of treatment on
gene expression. Using Affymetrix GeneChips®, we
have found that even in clinical remission, circulating immune cells from MS patients show profound
changes in gene expression compared to healthy
control subjects. Furthermore, we have identified a
gene expression pattern that appears to reflect the
activity of endogenous interferon-beta, and is associated with the expression of factors that control
disease activity in MS.
Gene expression is controlled by the activity of
transcription factors that are either constitutively
expressed in active forms, expressed in forms
that require activation by other signals, or are
inducible on cellular activation. The transcription
factors direct the expression of messenger RNA
(mRNA) from the genes encoded by the DNA
sequence, and after export from the nucleus the
mRNA provides a template for protein synthesis on ribosomes in the cytosol. Within the last
decade it has become apparent that endogenous,
small RNA molecules termed microRNA (miRNA)
have a key role in regulating the translation of
protein from mRNA in the cytosol. MiRNA can
either induce the degradation of mRNA, or can
inhibit the translation of mRNA into protein by
binding to the 3’ untranslated region of mRNA
molecules. Recent studies have identified miRNA
molecules that are differentially expressed in MS
patients and healthy controls, and it has been suggested that pathogenic immune activation in MS
may, indeed, reflect changes in miRNA expression.
We have studied miRNA expression in MS, and
have found that even patients with MS in clinical
remission show clear changes in the expression
of miRNA compared to healthy control subjects.
Furthermore, we have identified miRNA molecules
that are specifically regulated by treatment with
interferon-beta.
As miRNA have profound effects on the translation of protein within the cell, studies of miRNA
molecules that are differentially expressed in MS,
may provide the key to understanding the regulatory mechanisms involved in MS pathogenesis.
Indeed, as miRNA can now be therapeutically
introduced into specific human cells, treatment
with miRNA or with molecules blocking the effect
of individual molecules may by a feasible avenue
for the future treatment of MS. In ongoing studies, we are analysing the functional relevance of
the changes in miRNA expression observed in our
previous studies.
Biomarker studies
The term biomarker is used for, e.g., a gene product
or protein, that can be measured in blood or other
body fluids, and reflects a pathogenetically or therapeutically relevant in vivo process. Immunological
and molecular biology studies provide the platform
for the development of biomarkers, but the validation of these also requires access to collections of
well characterized samples from MS patients with
well characterized disease subtypes, who are fol-
lowed prospectively. Such biomarker studies are
important both for improving our understanding
of the pathogenesis of MS and for understanding
the therapeutic effects of immunomodulatory and
immunosuppressive treatment strategies.
Our group has been active in studying the immunological effects of treatment with interferon-beta.
These studies have resulted in the identification
of novel markers that may reflect the therapeutic
effect of treatment better than the currently used
biomarkers. Furthermore, we have used systematic
DNA array biomarkers studies to show that the
development of neutralizing anti-interferon-beta
antibodies results in complete abolishment of the
biologic effects of treatment. These studies have
also resulted in the identification of a gene expression pattern that appears to be associated with
disease protection, and may identify patients that
are more likely to remain in clinical remission on
treatment with interferon-beta.
Cerebrospinal fluid is well suited for studying
biomarkers that reflect pathogenic processes within
the intrathecal compartment. Although cerebrospinal fluid is more difficult to obtain on a routine
basis than blood samples, studies of such biomarkers have yielded important information about
the dynamics of immune cell infiltration, immune
regulation and tissue damage in MS. Most recently,
we have studied changes in cerebrospinal fluid in
patients treated with natalizumab (Tysabri), and
have identified an apparently central role for the
chemotactic factor (chemokine) CXCL13 in controlling the recruitment of immune cells to the brain
and spinal cord in MS.
The development of biomarkers that may serve
as surrogate outcomes in clinical trials is an important aim of the ongoing biomarker research at our
center. We assess changes in cerebrospinal fluid
biomarkers in patients with progressive MS that
participate in ongoing treatment trials, and analyse
the relationship between the different types of
biomarkers and clinical and magnetic resonance
imaging measures of disease activity and disease
severity. These studies provide the platform for
the design of future phase 2 studies of new MS
treatments that may be conducted more rapidly, requiring a lower number of patients than the current
protocols for phase 2 studies in MS.
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Research activities 2009
Neuropathology
Neuropathology research group:
Stephan Bramow, Per Soelberg Sørensen
(Henning Laursen, Laboratory of Neuropathology)
In close collaboration with Dr. Henning Laursen,
Laboratory of Neuropathology, Rigshospitalet and
Professor Hans Lassmann, Director of Institute
for Brain Research in Vienna, we have studied the
pathological correlates in brain and spinal cord of
patients with long-standing MS disease. We have
been able to show that inflammation and neurodegeneration are closely associated. We identified
a group of patients without any active or slowly
expanding demyelination in the brain, and these
patients did not show any signs of acute axonal
injury apart from confounding pathology unrelated
to MS such as Alzheimer disease.
Planimetric analysis of brain and spinal cords
from patients with clinically well characterized
primary progressive or secondary progressive MS
showed that at total demyelination was more profound in brains of secondary progressive patients
compared to primary progressive patients, whereas
demyelination in the spinal cord was similar. Active
demyelination correlated with shorter survival in
patients with secondary progressive MS.
We also demonstrated that remyelinated areas
had increased vulnerability and recurrent demye­
lination in the remyelinated areas were more
frequent than appearance of new demyelinating
plaques. This could explain why patients often
develop recurrence of symptoms in new relapses.
We also showed that diffuse inflammation in the
white-matter outside plaques correlated with the
load of active demyelination elsewhere in the brain.
The correlation between diffuse new inflammation and active focal demyelination emphasizes the
necessity of an action on both sides of the bloodbrain-barrier for new treatments of progressive MS.
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DANISH Multiple sclerosis CEN TER annual report 2009
Routine analysis in
neuroimmunology
laboratory
Neuroimmunology Laboratory
research group:
Poul Erik H. Jensen, laboratory technicians: Joy
Mendel-Hartvig, Marie Koefoed, Michael Kolbjørn
Jensen, Vibeke Fuglholt
Diagnostic evaluation
The presence in CSF of oligoclonal IgG-bands is of
interest in the diagnosis of Multiple Sclerosis (MS).
In 2009 we have analyzed 624 patient samples,
using isoelectric focusing of CSF and corresponding plasma samples for the characterization of IgG
bands.
In the autoimmune disease Myasthenia gravis
(MG), autoantibodies against the acetylcholine
receptor (AChR) may cause a diminished binding of
ACh on muscular surfaces and thereby a reduced
impulse transmission to the postsynaptic membrane
of the neuromuscular endplate occurs. For diagnostic and therapeutic purposes, we measure the
concentrations of these autoantibodies from patient
serum samples, using a radio-immunoassay kit, and
in year 2009 we analyzed 1233 patient samples.
Measurement of neutralizing antibodies
Subgroups of MS patients, treated with interferonbeta or Tysabri, generate neutralizing antibodies,
which diminish the therapeutic effects. Interferonbeta molecules bind to leucocytes and a specific
up-regulation of MxA mRNA in the cells occur.
Neutralizing antibodies may abolish this effect,
and therefore we measure the neutralization of
interferon-beta by antibodies in new cell-culture
assay based on luciferase-induced expression, and
further by the MxA mRNA-expression as a biological response to treatment with interferon-beta. In
2009 we have analyzed 244 patient samples for
neutralizing antibodies, and 131 patient samples
for MxA mRNA expression. Furthermore, we have
screened 36 patient samples for interferon-beta
binding antibodies using an ELISA-assay.
The action of Tysabri differs from interferonbeta, since it blocks mononuclear cell binding to
endothelial cells. In this way Tysabri inhibits mononuclear cells from entering the central nervous
system. The generation of neutralizing antibodies to
Tysabri in MS patients block the biological effects
of Tysabri. In 2009 we analyzed 634 blood samples
for the presence of antibodies to Tysabri by ELISA.
DANISH Multiple sclero sis CE N TER ann ual report 2009
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Scientific
publications,
prizes,
collaboration,
acknowledgements
Scientific publications 2008-2009
Prizes
Honorary offices
Scientific collaboration
Acknowledgements
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DANISH Multiple sclerosis CEN TER annual report 2009
DANISH Multiple sclero sis CE N TER ann ual report 2009
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Scientific publications
Publications 2008
Peer reviewed original papers
Krakauer M, Sørensen PS, Khademi M, Olsson T, Selle­
bjerg F. Increased IL-10 mRNA and IL-23 mRNA
expression in multiple sclerosis - interferon-beta
treatment increases IL-10 mRNA expression while
reducing IL-23 mRNA expression. Mult Scler 2008;14:
622-30.
Sellebjerg F, Datta P, Larsen J, Rieneck K, Alsing I,
Oturai A, Svejgaard A, Sørensen PS, Ryder LP. Gene
expression analysis of interferon-beta treatment in
multiple sclerosis. Mult Scler 2008;14: 615-21.
Hedegaard CJ, Krakauer M, Bendtzen K, Lund H,
Sellebjerg F, Nielsen CH. T helper cell type 1 (Th1),
Th2 and Th17 responses to myelin basic protein and
disease activity in multiple sclerosis. Immunology
2008;125:161-69.
International Multiple Sclerosis Genetics Consortium.
Refining novel associations in multiple sclerosis.
Lancet Neurol 2008; 7: 567-69.
Hedegaard CJ, Krakauer M, Bendtzen K, Sorensen PS,
Sellebjerg F, Nielsen CH. The effect of beta-interferon
therapy on myelin basic protein-elicited CD4+ T cell
proliferation and cytokine production in multiple
sclerosis. Clin Immunol 2008;129: 80-9.
Lorentzen Å, Smestad C, Lie B, Oturai A, Åkesson E,
Saarela J, Myhr K-M, Vartdal F, Celius E, Sørensen
PS, Hillert J, Spurkland A, Harbo HF. The SH2D2A
gene and susceptibility to multiple sclerosis. J Neuroimmunol 2008;197(2):152-8.
International Multiple Sclerosis Genetic Consortium
(IMSGC): Booth D, Heard R, Stewart G, Goris A,
Dobosi R, Dubois B, Oturai A, Soendergaard HB,
Sellebjerg F, Saarela J, Leppä V, Palotie A, Peltonen
L, Fontaine B, Cournu-Rebeix I, Clerget-Darpoux F,
Babron MC, Weber F, Holsboer F, Müller-Myhsok B,
Rieckmann P, Kroner A, Graham C, Vandenbroeck
K, Hawkins S, D’Alfonso S, Bergamaschi L, Naldi P,
Guerini FR, Salvetti M, Galimberti D, Hintzen R, van
Duijn C, Lorentzen AR, Celius EG, Harbo HF, Spurkland A, Cucca F, Marrosu MG, Comabella M, Montalban X, Villoslada P, Olsson T, Kockum I, Hillert J,
Ban M, Walton A, Sawcer S, Compston A, Hawkins C,
Mihalova T, Robertson N, Ingram G, Hafler DA, Rioux
J, Daly M, Barcellos L, Ivinson A, Pericak-Vance
M, Oksenberg J, Hauser SL, McCauley J, Sexton D,
Haines J. Refining novel associations in multiple
sclerosis. Lancet Neurol 2008;7(7):567-9.
26
DANISH Multiple sclerosis CEN TER annual report 2009
Sorensen PS. Intravenous polyclonal human immunoglobulins in multiple sclerosis. Neurodegener Dis
2008;5(1):8-15.
Sorensen PS, Koch-Henriksen N, Flachs EM, Bendtzen
K. Is the treatment effect of IFN-beta restored after
the disappearance of neutralizing antibodies? Mult
Scler 2008;14(6):837-42.
Bramow S, Faber-Rod JC, Jacobsen C, Kutzelnigg A,
Patrikios P, Sorensen PS, Lassmann H, Laursen H.
Fatal neurogenic pulmonary edema in a patient
with progressive multiple sclerosis. Mult Scler
2008;14(5):711-5.
Fazekas F, Lublin FD, Li D, Freedman MS, Hartung HP,
Rieckmann P, Sørensen PS, Maas-Enriquez M, Sommerauer B, Hanna K; PRIVIG Study Group; UBC MS/
MRI Research Group. Intravenous immunoglobulin in
relapsing-remitting multiple sclerosis: a dose-finding
trial. Neurology 2008;71(4):265-71.
Giovannoni G, Barbarash O, Casset-Semanaz F, King J,
Metz L, Pardo G, Simsarian J, Sørensen PS, Stubinski
B. Safety and immunogenicity of a new formulation
of interferon beta-1a (Rebif(R) New Formulation) in
a Phase IIIb study in patients with relapsing multiple
sclerosis: 96-week results. Mult Scler 2008.
Elovaara I, Apostolski S, van Doorn P, Gilhus NE, Hietaharju A, Honkaniemi J, van Schaik IN, Scolding N,
Sørensen PS, Udd B. EFNS guidelines for the use of
intravenous immunoglobulin in treatment of neurological diseases: EFNS task force on the use of intravenous immunoglobulin in treatment of neurological
diseases. Eur J Neurol 2008;15(9):893-908.
Ravnborg M, Bendtzen K, Christensen O, Jensen P,
Hesse D, Tovey M, Sørensen PS. Treatment with
azathioprine and cyclic methylprednisolone has little
or no effect on bioactivity in anti-interferon beta
antibody-positive patients with multiple sclerosis.
Mult Scler 2008.
Other publications
Sellebjerg F, Oturai A, Sørensen PS; Dansk Neurologisk
Selskab. Multipel sklerose – genernes betydning.
Ugeskr Læger 2008;170: 1046.
Sørensen PS, Sellebjerg F. Biologisk behandling af multipel sklerose. Ugeskr Læger 2008;170:2156-9.
2008
2009
Mathiesen HK, Ravnborg M, Sørensen PS. Magnetic
resonance imaging in the diagnosis and follow-up of
multiple sclerosis. Ugeskr Laeger 2008;170(34):2579-81.
Sorensen PS, Sellebjerg F. Oral fumarate for relapsing-remitting multiple sclerosis. Lancet 2008;372(9648):14478.
Sørensen PS. REGARD: what can we learn from randomised, open-label, head-to-head studies? Lancet
Neurol 2008;7(10):864-6.
Sorensen PS. Can we spot the IFN-beta nonresponders?
Neurology 2008;71(24):1936-7.
Publications 2009
Peer reviewed original papers
Kemppinen A, Suvela M, Tienari PJ, Elovaara I, Koivisto
K, Pirttilä T, Reunanen M, Baumann P, Hillert J,
Lundmark F, Oturai A, Ryder L1, Harbo H, Celius
EG, Palotie A, Daly M, Peltonen L, Saarela J. MYO9B
polymorphisms in multiple sclerosis. Eur J Hum Genet
2009;17(6):840-3.
Khademi M, Börnsen L, Rafatnia F, Andersson M, Brundin
L, Piehl F, Sellebjerg F, Olsson T. The effects of natalizumab on inflammatory mediators in multiple sclerosis:
prospects for treatment-sensitive biomarkers. Eur J
Neurol 2009;16: 528-36.
Sellebjerg F, Börnsen L, Khademi M, Krakauer M, Olsson T, Frederiksen JL, Sorensen PS. B cell chemokine
CXCL13 in cerebrospinal fluid in active MS. Neurology
2009;73: 2003-10.
Mero I-L, ÅR Lorentzen, Ban M, Smestad C, Celius EG,
Aarseth J, Myhr K-M, Link J, Hillert J, Olsson T, Kockum I, Masterman T, Oturai AB, Søndergaard HB, Sellebjerg F, Saarela J, Kemppinen A, Elovaara I, Spurkland
A, Dudbridge F, Lie BA, Harbo HF. A rare variant of the
TYK2 gene is confirmed to be associated with multiple
sclerosis. Eur J Hum Genet 2010;18(4):502-4.
Sellebjerg F, Börnsen L, Khademi M, Krakauer M, Olsson
T, Frederiksen JL, Sørensen PS. Increased cerebrospinal
fluid concentrations of the chemokine CXCL13 in active
MS. Neurology 2009;73(23):2003-10.
Ockinger J, Stridh P, Beyeen AD, Lundmark F, Seddighzadeh M, Oturai A, Sørensen PS, Lorentzen AR, Celius EG,
Leppä V, Koivisto K, Tienari PJ, Alfredsson L, Padyukov
L, Hillert J, Kockum I, Jagodic M, Olsson T. Genetic
variants of CC chemokine genes in experimental autoimmune encephalomyelitis, multiple sclerosis and
rheumatoid arthritis. Genes Immun 2010;11(2):142-54.
Hesse D, Sellebjerg F, Sorensen PS. Absence of MxA
induction by interferon beta in patients with MS
reflects complete loss of bioactivity. Neurology
2009;73(5):372-7.
Sellebjerg F, Krakauer M, Hesse D, Ryder LP, Alsing I,
Jensen PE, Koch-Henriksen N, Svejgaard A, Soelberg
Sørensen P. Identification of new sensitive biomarkers
for the in vivo response to interferon-beta treatment
in multiple sclerosis using DNA-array evaluation. Eur J
Neurol 2009;16(12):1291-8.
Enevold C, Oturai AB, Sørensen PS, Ryder LP, KochHenriksen N, Bendtzen K. Multiple sclerosis and
polymorphisms of innate pattern recognition receptors
TLR1-10, NOD1-2, DDX58, and IFIH1. J Neuroimmunol 2009;212(1-2):125-31.
Sorensen PS, Mellgren SI, Svenningsson A, Elovaara I,
Frederiksen JL, Beiske AG, Myhr KM, Søgaard LV,
Olsen IC, Sandberg-Wollheim M. NORdic trial of oral
Methylprednisolone as add-on therapy to Interferon
beta-1a for treatment of relapsing-remitting Multiple
Sclerosis (NORMIMS study): a randomised, placebocontrolled trial. Lancet Neurol 2009:519-29.
Oturai AB, Koch-Henriksen N, Petersen T, Jensen PE,
Sellebjerg F, Sorensen PS. Efficacy of natalizumab
in multiple sclerosis patients with high disease
activity: a Danish nationwide study. Eur J Neurol
2009;16(3):420-3.
Frischer JM, Bramow S, Dal-Bianco A, Lucchinetti CF,
Rauschka H, Schmidbauer M, Laursen H, Sorensen PS,
Lassmann H. The relation between inflammation and
neurodegeneration in multiple sclerosis brains. Brain
2009;132(Pt5):1175-89.
Koch-Henriksen N, Sorensen PS, Bendtzen K, Flachs EM.
The clinical effect of neutralizing antibodies against
interferon-beta is independent of the type of inter-
DANISH Multiple sclero sis CE N TER ann ual report 2009
27
Scientific publications
feron-beta used for patients with relapsing-remitting
multiple sclerosis. Mult Scler 2009;15(5):601-5.
Hedegaard CJ, Chen N, Sellebjerg F, Sørensen PS, Leslie
RG, Bendtzen K, Nielsen CH. Autoantibodies to myelin basic protein (MBP) in healthy individuals and in
patients with multiple sclerosis: a role in regulating
cytokine responses to MBP. Immunology 2009;128(1
Suppl):e451-61.
Nielsen TR, Rostgaard K, Askling J, Steffensen R,
Oturai A, Jersild C, Koch-Henriksen N, Sørensen
PS, Hjalgrim H. Effects of infectious mononucleosis
and HLA-DRB1*15 in multiple sclerosis. Mult Scler
2009;15(4):431-6.
O’Connor P, Comi G, Montalban X, Antel J, Radue EW,
de Vera A, Pohlmann H, Kappos L; FTY720 D2201
Study Group. Oral fingolimod (FTY720) in multiple
sclerosis: two-year results of a phase II extension
study. Neurology 2009;72(1):73-9.
Hesse D, Frederiksen JL, Koch-Henriksen N, Schreiber
K, Stenager E, Heltberg A, Ravnborg M, Bendtzen
K, Sellebjerg F, Sorensen PS. Methylprednisolone
does not restore biological response in multiple
sclerosis patients with neutralizing antibodies against
interferon-beta. Eur J Neurol 2009;16(1):43-7.
Ravnborg M, Bendtzen K, Christensen O, Jensen PE,
Hesse D, Tovey MG, Sørensen PS. Treatment with
azathioprine and cyclic methylprednisolone has little
or no effect on bioactivity in anti-interferon beta
antibody-positive patients with multiple sclerosis.
Mult Scler 2009;15(3):323-8.
Giovannoni G, Barbarash O, Casset-Semanaz F, King J,
Metz L, Pardo G, Simsarian J, Sørensen PS, Stubinski
B; Rebif New Formulation Study Group. Safety and
immunogenicity of a new formulation of interferon
beta-1a (Rebif New Formulation) in a Phase IIIb
study in patients with relapsing multiple sclerosis:
96-week results. Mult Scler 2009;15(2):219-28.
Jagodic M, Colacios C, Nohra R, Dejean AS, Beyeen AD,
Khademi M, Casemayou A, Lamouroux L, Duthoit
C, Papapietro O, Sjöholm L, Bernard I, Lagrange D,
Dahlman I, Lundmark F, Oturai AB, Soendergaard
HB, Kemppinen A, Saarela J, Tienari PJ, Harbo HF,
Spurkland A, Ramagopalan SV, Sadovnick DA, Ebers
GC, Seddighzadeh M, Klareskog L, Alfredsson L,
Padyukov L, Hillert J, Clanet M, Edan G, Fontaine
B, Fournié GJ, Kockum I, Saoudi A, Olsson T. A role
for VAV1 in experimental autoimmune encephalomyelitis and multiple sclerosis. Sci Transl Med
2009;1(10):10ra21.
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DANISH Multiple sclerosis CEN TER annual report 2009
2009
Other publications
Waldemar G, Boysen GM, Høgenhaven H, Knudsen GM,
Sørensen PS, Vissing J. Neurologi. I: Schaffalitzky de
Muckadell OB; Haunsø S, Vilstrup H (red.). Medicinsk
Kompendium 17. Udgave, Nyt Nordisk Forlag Arnold
Busck, København 2009; pp.2418-2623.
Sorensen PS. How effective is natalizumab as secondline treatment for multiple sclerosis in daily clinical
praxis? Eur J Neurol 2009;16(3):287-8.
Sorensen PS. Management of patients with neutralizing
antibodies against interferon-beta: stop IFN-beta
therapy or wait for the antibodies to go away? Eur J
Neurol 2009;16(1):1-2.
Prizes and honorary offices
Prizes
Annette Oturai
“Henry Hansen and wifes grant”: 400.000 kr, donated for many years of scientific working
within multiple sclerosis, 2009.
Honorary offices
Annette Bang Oturai has held the following honorary offices:
A: National
Board member of the Danish Sociaty for Multiple Sclerosis (DAREMUS)
Finn Sellebjerg has held the following honorary offices:
A: National
Chairman of the Danish Society for Research in Multiple Sclerosis (MS) and chairman of the
research board of the Danish MS Society.
B: International
Chairman of the task force on treatment of MS relapses and member of the scientist panel
on demyelinating disease of the European Federation of Neurological Societies.
Per Soelberg Sorensen has held the following honorary offices:
A: National
Chairman of the Foundation for Research in Neurology, 1986Chairman of the Scientific Advisory Committee of the Danish MS Society, 2004-2010
Chairman of the Danish Multiple Sclerosis Group, 1996B: International
Executive Board Member of the European Charcot Foundation for Research in Multiple
Sclerosis, 1994Member, Medical Advisory Board of the International Federation of Multiple Sclerosis
Societies, 1998Member, US National Multiple Sclerosis Society Advisory Committee on Clinical Trials, 2000Chairman, Scientist Panel on Multiple Sclerosis, European Federation of Neurological
Societies, 2003Chairman, Publication Committee, European Federation of Neurological Societies, 2003Executive Board Member of the European Committee for Treatment and Research in
Multiple Sclerosis (ECTRIMS), 2009-
DANISH Multiple sclero sis CE N TER ann ual report 2009
29
Scientific collaboration
Scientific collaboration
National
Danish Research Centre for Magnetic Resonance,
Copenhagen University Hospital, Hvidovre, Copenhagen, Denmark
Department of Clinical Neuroscience and Centrum
for Molecular Medicine Karolinska Insitutet at
Karolinska University Hospial, Solna 171 76 Stockholm,
Sweden (Professor Tomas Olsson, associate Professor
Ingrid Kockum)
The Danish Multiple Sclerosis Register, Copenhagen
University Hospital, Rigshospitalet, Copenhagen, Denmark (Nils Koch-Henriksen, MD)
Department of Neurology, Lund University Hospital,
Lund, Sweden (Professor Magnhild Sandberg-Wollheim)
Department of Clinical Immunology, Copenhagen
University Hospital, Rigshospitalet, Denmark (Jacob
Larsen MD, Lars Ryder , Klaus Rieneck, MD, Hans O.
Madsen, MD)
Department of Neurology, Gothenburg University Hospital, Gothenburg, Sweden (Professor Oluf Andersen)
Institute for Inflammatory Research, Copenhagen
University Hospital, Rigshospitalet, Denmark (Christian
Enevold-Johansen MD, Professor Klaus Bendtzen)
Department of Epidemiology Research, Statens Serum
Institut, Copenhagen, Denmark (Trine Rasmussen
Nielsen, MD, Henrik Hjalgrim, MD, professor Mads
Melbye, Peter Michael Bager, ph.d.)
Department of Human Genetics, Aarhus University,
Denmark (Bjørn Andersen Nexø)
Laboratory of Neuropathology, Copenhagen University
Hospital Rigshospitalet, Copenhagen, Denmark (Henning Laursen, MD)
Institute for Molecular Medicine Finland, University
of Helsinki, Finland (Janna Saarela)
University of Cambridge, Neurology Unit, Addenbrooke’s Hospital, Cambridge, United Kingdom
(Stephen Sawcer, MD, Professor Alastair Compston)
“International Multiple Sclerosis Genetic Consortium” (IMSG): collaboration between 20 countries from
Europe, USA, Canada and Australia
Department of Immunopathology, Brain Research
Center, Medical University of Vienna, Vienna, Austria.
(Professor Hans Lassmann and Josa Frischer, MD)
Department of Neurology, Mayo Clinic, Rochester,
Minnesota, USA (Professor Claudia F. Lucchinetti)
VU Medical Centre, Amsterdam, The Netherlands
(Professor Chris Polman)
International
Nordic MS Genetic Network: Collaboration between the
Nordic countries: Sweden (Huddinge, Lund, Gothenburg, Stockholm), Norway (Oslo, Bergen), Finland
(Helsinki) and Denmark (Copenhagen)
Institute of Immunology, Rikshospitalet, University
Hospital, Oslo, Norway (Anne Spurkland, MD, Hanne F.
Harbo, MD, professor Frode Vartdal)
Department of Neurology, Ullevål University Hospital,
Oslo, Norway (Elisabeth G Celius, MD)
Department of Neurology, Haukeland Hospital, Bergen,
Norway (Professor Kjell-Morten Myhr)
Department of Neurology, Huddinge University Hospital, Karolinska Institute, Huddinge, Sweden (Professor
Jan Hillert, Eva Åkesson, MD, Helena Modin, MD)
30
DANISH Multiple sclerosis CEN TER annual report 2009
Utrecht University, Utrecht, The Netherlands (Professor
Hub Schellekens)
Heinrich-Heine-University, Düsseldorf, Germany
(Professor Hans-Peter Hartung)
Ospedale Universitario San Luigi, Torino, Italy
(Professor Auturio Bertolotto)
Queen Square, London, The United Kingdom (Professor
Gavin Giovannoni)
Innsbruck Medical University, Innsbruck, Austria
(Professor Florian Deisenhammer)
General Charles University, Prague, Czech Republic
(Professor Eva Havrdova)
Collaboration with
pharmaceutical companies
on clinical trials
Novartis, Denmark
Merck Serono Nordic, Denmark, Norway and Sweden
Biogen idec, Denmark and USA
Teva/Aventis, Israel and Denmark
Sanofi-aventis, Denmark
Bayer Schering, Germany
Octapharma, Austria
Genmab, Denmark
Genzymes, Holland
BioMS
Glaxo Smith-Kline
Acknowledgements
Danish Multiple Sclerosis Research Center has received
generous support from a number of public and private
research funds:
Danish Medical Research Council
Danish MS Society
Warwara Larsen Foundation
Rigshospitalets Scientific Board
EU Sixth Framework Programme
Brdr. Røjne Holding
Jeppe Juel Memorial Legacy
RoFar Foundation
Roche Denmark
The Danish Strategic Research Council
The Johnsen Memorial Foundation
The Lounkær Foundation
Biogen idec
Sanofi-aventis
Merck Serono
Ejner Jonasson og Hustrus mindelegat
Fondsbørsvekselerer Henry Hansen og Hustrus Legat
DANISH Multiple sclero sis CE N TER ann ual report 2009
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DANISH Multiple sclerosis CEN TER annual report 2009