1. No category

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Lymphokine
Induction
Melchor
By
of NK-Like
Alvarez
de Mon.
Manuel
0.
Cytotoxicity
Juan
Casas,
T cells
from
patients
with
B cell chronic
lymphocytic
leukemia
(B-CLL) exhibit
defective
natural
killer (NK) activity. In this study,
we have analyzed
the cytotoxic-inducer
effects
of gamma
interferon
(‘y-IFN)
and
supernatants
containing
interleukin
2 (II 2 sup).
T cells from
patients
with
B-CLL
were
incubated
with
‘y-IFN
or IL 2 sup.
“y-IFN
in a 1 5,000-
B
CELL
to 20,000-dalton
CHRONIC
molecular
lymphocytic
characterized
by the
mulation
of B cells, which
leukemia
monoclonal
are arrested
tant;
did
these
patients
also show functional
malities.2’3
In addition,
it has been
from B-CLL
cytotoxicity
against
poiesis
It has
been
demonstrated
their
ability
without
prior
and
control
that
classic
2 (IL
There
2)89
tioned
medium.
2 supernatants
This study examined
(IL 2 sup) on T cells
tion
with
NK
normal
NK
hemato-
cells,
defined
IL
reports
describing
the in vitro
from B-CLL
activity
against
that short-term
2 sup
can
T cells
from
induce
strong
B-CLL
by
targets
(IFN)
cells’#{176}” that
lyse tumor
cell
culture
with
lymphokine-condi-
of cytotoxic
demonstrate
unstimulated
present
in these
of this disease,
in the surveillance
are also
of killer
long-term
the induction
Our results
natural
cellular
to lyse a number
of tumor
cell line
activation,
are modulated
by interferon
interleukin
that
from
and phenotypic
abnorshown
that
E rosette-
of both
the induction
targets
during
is
and accudifferentia-
patients
mediate
defective
and antibody-dependent
tumors
and in the
and viral infection.6’7
line
effect of IL
patients
in
tumor
target
cells.
hours)
incuba-
(18
NK-like
patients
activity
that
lack
in
classical
activity.
AND
MATERIALS
METHODS
Patient
population.
Twenty-six
previously
untreated
patients
with clinically,
histologically,
and immunologically
defined B-CLL
(according
to the Chronic
Leukemia
Myeloma
Task Force 1973)12
From
the
.
Supported
Fondo
of
de Hierro,
by Grants
Address
March
reprint
Interna,
Internal
No.
369/81,
18, 1985;
requests
Clmnica
135/82,
and
accepted
July
Alvarez
de
Hierro,
Immunology,
Madrid.
0671/83from
Social,
to Dr M.
Puerta
and
Autbnoma,
de la Seguridad
Porres, 4, 28035 Madrid,
Spain.
(e I 986 by Grune & Stratton,
Inc.
0006-4971/86/6701-0036$03.OO/O
228
Medicine
Universidad
de Investigaciones
Submitted
Medicina
Departments
Puerta
the
Spain.
26,
1985.
de Mon,
ci
San
Toribio,
by gel
filtration
target
cells;
of this
NK-sensitive
chromatography
directed
was
against
(c) was
not
correlated
with
supernaand
NK-
the
basal
levels
of NK activity;
and (d) was not associated
with
a
development
or augmentation
of the proportion
of lymphocytes
with
classic
NK cell
phenotype.
Taken
together.
these
results
demonstrate
that
unstimulated
T cells from
B-CLL
but
patients.
not
lack
incubated
‘y-IFN,
have
of classic
a 1986
NK
by Grune
strong
briefly
(1 8 hours)
NK-like
cytotoxicity.
with
IL 2 sup
despite
the
activity.
& Stratton,
Inc.
were studied.
Their median
age was 68 ± 9. Controls
were 14
healthy, age- and sex-matched
individuals.
Culture
medium.
RPMI
1640 (Microbiological
Assoc, Walkersville, Md) supplemented
with 1% L-glutamine
(Flow Lab, Irvine,
UK), 0.5% HEPES
(Flow Lab), and 1% penicillin
streptomycin
(Difco Lab, Detroit) was used for cultures.
This will be referred to as
complete
medium.
Isolation
oflymphoid
cells.
Peripheral
blood mononuclear
cells
(PBMs)
were obtained
from the heparinized
venous blood of the
subjects
by Ficoll-Hypaque
gradient
centrifugation.
T lymphocytes
were separated
by their capacity
to form stable
rosettes
with
neuraminidase-treated
sheep red blood cells (SRBCs).
Briefly, ESRBCs
(T cells) were obtained
after incubation
of PBMs
with
neuraminidase-treated
SRBCs for two hours at 4 #{176}C
and centrifugation on a Ficoll-Hypaque
gradient.
The interphase,
containing
mainly B cells, null cells, and macrophages,
is referred
to as non-T
cells. A Tcell purity of9O% to94%(51%
± 6% OKT4,
41% ± 10%
OKT8)
in the control group and 78% to 94% (36% ± 5% OKT4,
34% ± 4% OKT8)
in the B-CLL
group
was obtained
after
subjecting
pellets to hypotonic
lysis in order to eliminate
SRBCs.
Production
ofIL
2 supernatants.
IL 2 sups were obtained
from
human tonsil lymphocytes
stimulated
with 1% phytohemagglutinin
(PHA)
(Difco Lab) for 24 hours, as previously
described.’3
The
culture supernatants
were concentrated
by double-step
precipitation
with (NH4)2S04
and dialyzed
against
phosphate-buffered
saline
(PBS), pH 7.4. The remaining
PHA was removed
by chicken
red
blood cell absorption,
as reported elsewhere.’4
Partially
purified IL 2
was obtained
by gel filtration
chromatography.’5
The IL 2 activity
was recovered
in fractions corresponding
to a molecular
weight (mol
wt) of 15,000 to 20,000 daltons.
IL 2 was also obtained
from a tumor T cell line. The Gibbon
lymphosarcoma
cell line, MLA-144,
provided
by Dr H. Rabin
(Frederick
Cancer Research
Center, Frederick,
Md), spontaneously
releases
a factor that is biologically
and biochemically
similar to
human
IL 2.16 MLA-I44
cells were cultured
at 2 x 106 cells per
milliliter
in complete
medium without serum for 48 hours. After the
culture
Clmnica
Luisa
B-.CLL
fraction
are indications
but that T cells
cytotoxicity.4’5
The defective
NK activity
patients
may play a role in the development
since NK activity
is thought
to be involved
Maria
From
Dur#{233}ntez
(b)
resistant
(B-CLL)
expansion
in their
tion pathway
at an early
stage.’
There
not only B cells are affected
in B-CLL,
positive
cells
killer
(NK)
weight
Laguna,
Alberto
obtained
not modulate
the very low or undetectable
levels
of NK
activity
present
in the T cell population.
However.
the IL 2
sup induced
a potent
cellular
cytotoxicity
against
NKsensitive
and NK-resistant
tumoral
target
cells.
This cytotoxic
inducer
effect
(a) was present
in lectin-free
IL 2 sup
and
Rafael
de Land#{233}zuri, and
in T Cells
Servicio
de
Martin
de
period,
supernatants
were
collected
and
concentrated
as
previously
described.
To study the production
of IL 2 by the T cells from either patients
or controls,
5 x 106 cells from each subject
and control
were
resuspended
in 1 mL of complete
medium
supplemented
with I %
fetal bovine serum (FBS) and cultured
for 24 h in the presence of I %
PHA in round-bottom
culture tubes (Falcon, Oxnard,
Calif). At the
end of the culture period, the supernatants
were collected
and stored
at - 20 #{176}C
until tested. IL 2 assays were performed
as previously
‘7
PHA-stimulated
L ofcomplete
cultures.
medium
Blood,
Purified
with
T cells
were
10% FBS and 1% PHA,
Vol 67. No
1 (January),
cultured
in 200
at a density
1986:
pp 228=232
of
From www.bloodjournal.org by guest on January 26, 2015. For personal use only.
NK-LIKE
ACTIVITY
IN
T CELLS
FROM
229
B-CLL
1.5 x i0 cells per well, in triplicates
in 96-well plates (Dynatech,
Billingshurst,
UK) for three days. Plates were incubated
at 37 #{176}C
in
a 5% CO2 humid atmosphere.
On day 2, 1 Ci of [3H]-thymidine
was added, and 24 hours later, cultures
were harvested
(Skatron,
AS.,
Lyerbyen,
Norway)
and counted
in a liquid scintillation
counter.
Cell
cultures
with
IL 2 sup.
Cell cultures
were set up in
macrowells
(1.6 x 106 cells per milliliter)
in complete
medium
supplemented
with 10% FBS and in the presence
or absence
of
different
concentrations
of IL 2 sup. The cultures
were carried out
for variable periods.
Cells
cultured
with
‘y-IFN.
As previously
described,
effector
cell populations
were cultured
in macrowells
(1.6 x 106 cells per
milliliter)
in complete
medium supplemented
with 10% FBS for 18
hours. In the last two hours, we added
I ,000 IU/mL
of human
‘y-IFN
(specific activity 2 x l0 lU/mg
protein HEM)
(Research
Inc. Rockville,
Md).
Cytotoxicity
assays.
Cytotoxicity
was quantified
by a 5’Crspecific release assay using the following tumor target cells: K-562,
Chang, MI- 1 , Alab, Raji, Molt-4, Molt-4-Thioguanin
resistant,
and
iurkat.
Assays were performed
at effector-target
ratios of 50, 25,
and 12:1 in 96-microwell,
round-bottom
plates.
After
a fourincubation,
100 L of supernatant
from each well was collected
and
analyzed
in a gamma counter. The specific lysis (SL) and lytic units
(LU) were calculated
as previously
Cell surface
markers.
Indirect
immunofluorescence
(lIF) with
the monoclonal
antibodies
(OKT4, OKT8 [Ortho, Raritan,
Ni], and
Leu 1 lb [Becton Dickinson,
Mountain
View, Calif] was performed
as described.’9
Samples
were prepared
as described
for IIF’9 and
analyzed
with an EPICS-C
Flow Cytometer
(Coulter
Electronics,
Hialeah,
Fla).
j
r-
Ei
-t
-J
4o
0
EFECTOR/TARGET
ceLL
RATIO
Fig 1 .
NK activity
of T cells from B-CLL patients
and normal
controls
at different
effector-target
cell ratio. The thick horizontal
segments
represent
the geometric
medium
of each population.
The shaded areas represent
two
standard
deviations
of the mean
of each population.
The extremes
of the rectangles
represent
the
rank of distribution
of the data for each population.
K-562
cells
were used as target
cells. .
Controls;
.
B-CLL.
RESULTS
activity
NK
non-T
cells
from
I 3 normal
As can
B-CLL
lack
a significantly
of
population
contaminating
observed
CLL
NK
was
cells
present
experiments
did not inhibit
(data
in
not due to inhibition
non-T
cells
(range,
NK
ofthe
by
and
the
‘y-IFN
investigated
K-562
capacity
20
that
Therefore,
respect
cells.
was
at time
x
LU
T
non-T
the cytotoxic
when
cells
of
we
from
activity
activity
2 sup.
B-
<
IL
against
4 LU
to ‘y-IFN,
x
10_6
cells;
used.
cytotoxic-inducer
T cells
from
K-562
cytotoxic
(P < .05,
of T cells
signed-ranked
activity
Wilcoxon
B-CLL
patients
(patients
I, II,
patients).
When
together,
the
1). After
incubation
with
was a significant
increase
were
IV,
and
the
all the normal
test).
In contrast,
either
or
unresponsive
minimally
of all
increments
treatment
were
Wilcoxon
signed-ranked
in
VI)
data
observed
in
40
this
effect
12 cases
two
hours
LU
x lO_6).
period
of incuof
of B-CLL
IL
2 with
with
were
test).
ranked
=
very
the
levels
and
the
NK
were
.05,
values
after
(P > .05,
when all
examined,
we
least squares
observed
of
the
analyzed
activity
of cytotoxicity
basal
of
were
after
NK
activity
(Fig 2).
The incubation
with IL 2 sup of T cell from each of the
B-CLL
patients
induced
a significant
increase
in their ability
to lyse K-562
target
cells, which
proved
significant
when all
the cases
in their
lytic
for I 8 hours
after
between
treatment
‘y-IFN
controls
T cells
(rest
significant
other
hand,
<
1,000
in the
to ‘y-IFN
so
12 patients
not statistically
test).
On the
‘y-IFN
method)
target
To this end, the T
with IL 2 sup, and
was tested
against
an increase
incubation
10_6; and
after
18 hours,
in the following
experiments,
was routinely
compare
the
(Table
there
the data from either
patients
or controls
observe
a significant
correlation
(P
of normal
In preliminary
experiments,
2 sup could
induce
cytotoxic
This revealed
maximal
after
0
these
lymphokines
IU/mL
of “y-IFN,
from
cell-enriched
levels of NK activity
and T cells from controls
NK activity
were incubated
for 18 hours
with
did
cytotoxic
IL
whether
target
that
units
bation
To
activity
by the small number
6% to 22%),
since
activity
in T cells from B-CLL
patients.
cells were incubated
for two or I 8 hours
subsequently,
their
cytotoxic
activity
(lytic
NK activicells from
not shown).
Modulation
we
low or absent
with normal
PBMs,
T, and
B-CLL
and from
cells.
untreated
depressed
activity
in mixing
patients
cells
non-T
with
with normal
controls
(P < .0001,
Mann-Whitney
Neither
PBMs
nor non-T
cells
from
B-CLL
any significant
cytotoxic
activity
(data not shown).
mediated
The
26 patients
showed
compared
U test).
T, and
controls
were separated
to study their
be observed
in Fig I , E rosette-positive
ty.
T
ofPBMs,
cytotoxicity
together
observed
that
the
by IL 2 sup
were
obtained
induced
also
(P
.0 1 , Wilcoxon
analyzed
We
<
highest
signedlevels
precisely
of
in
those
patients
who
did not show
spontaneous
cytotoxic
activity
and did not respond
to ‘y-IFN.
We did not observe
any correlation,
either
in B-CLL
patients
or in normal
controls,
between
the basal
levels
of NK activity
and the
killing
activity
after
incubation
with
IL 2 sup
(Fig
2).
From www.bloodjournal.org by guest on January 26, 2015. For personal use only.
ALVAREZ
230
Table
1 . Effects
of ‘y-IFN
and IL 2 Sup on C ytotoxic
Activity
of T Cells
B-CLL
Against
K-562
Target
Cells
With
-y-IFN
After
Treatment
IL 2 Sup
With
Subjects
Patients
50/1
25/1
I
13
3
II
12
8
Ill
10
5
2
IV
5
2
0
V
3
0
0
VI
3
1
0
VII
12/1
ET AL
Lysis
Specific
Treatment
After
Basal
.
From
MON
DE
50/1
25/1
12/1
50/1
25/1
0
14
9
1
35
14
5
24
18
7
6
2
4
12/1
5
13
34
24
16
3
0
70
60
49
4
0
70
50
26
0
0
47
34
19
2
1
76
60
46
4
3
0
14
3
1
35
14
9
VIII
19
12
0
44
21
11
49
35
15
lx
13
9
1
19
13
11
43
23
19
x
17
9
4
29
22
16
51
38
26
Xl
14
10
6
34
27
14
49
42
23
XII
19
16
11
30
22
15
50
42
31
18±13
12±9
Patients
(±s)
11±6
6±4
2±3
50±14
6±6
36±15
23±13
Controls
(i±s)
T cells
hours,
45±18
from
(1
y-IFN
analyzed
for
1 2 B-CLL
.000
cytotoxic
36±15
patients
and
lU/mL)
being
activity
against
25±17
six normal
added
K-562
in the
72±15
controls
were
last
hours;
target
two
cells.
The
incubated
data
61±9
for
or complete
are given
45±22
1 8 hours
either
medium
plus
with
20%
as the percentage
76±20
complete
IL 2 sup.
of specific
66±25
medium;
After
complete
this
51±26
medium
period
of time,
lysis at effector-target
for
the
cell ratios
the first
cultures
16
were
of 50: 1 , 25:1,
12:1.
and
This finding
led us to study the possibility
that the T cells
from B-CLL
patients
could be deficient
in their synthesis
of
IL 2. We
B-GIL
ThTROIS
(A-I)
cells
(B-I)
found
from
that
nine
IL 2 production
patients
were
by PHA-stimulated
similar
to that
controls
(Fig 3). However,
the proliferative
stimulation
of the T cells
from
these
S
T
of six normal
response
B-CLL
to PHA
patients
S
CPM
V.
(K53)
0810010
r’OM8e
S
0
S
0
S
0
8-CLL
o
S
#{149}
S
(B-2)
WNTROLS
(A-2)
#{149}
5
0
I2
c0
SS.
S
0S
5
0
5
0
0
S
:
0
S
S
N
0
0S
,. .O314
F,
0O3
S
40
S
S
S
S
0
8s
S
a
40
ZJ
40
LYSIS
#{149}1.
ECIFIC
(8ASt
40
60
813
LEVELS)
V2
Fig 2.
Correlations
between
the basal levels of cytotoxicity
and (A) the lytic activity
of T cells from B-CLL after incubation
with
‘y-IFN
(A-i ) or IL 2 sup (A-2) and (B) the lytic activity
of T cells from
controls
after incubation
with ‘y-IFN (B-i)
of IL 2 sup (8-2).
The
cytotoxic
against
activity
K-562
is expressed
target
cells
as
a percentage
at an effector-target
of
specific
cell ratio
lysis
of 50:i.
‘/5
L-2
.JP
DILUTiONS
Fig 3.
Proliferative
response
of long-term
cultured
human
T
after a 36-hour
incubation
period.
in culture.
with
different
proportions
of IL 2 sup produced
by T cells from either
B-CLL
patients
(#{149})
or normal controls
(0).
cells
From www.bloodjournal.org by guest on January 26, 2015. For personal use only.
NK-LIKE
ACTIVITY
(34,407
IN
15,709
±
decreased
with that
(P
of
T CELLS
FROM
cpm/I.5
231
B-CLL
x i0
cells)
.01, Mann-Whitney
the normal
controls
was
U test)
(87,082
<
significantly
_______________
C14660
when compared
± 21,179
cpm/
MT-I
1.5 x 10 cells).
ALAS
We also ruled
cytotoxic
activity
out the possibility
that
by IL 2 sup was due
contaminating
non-T
cells,
since
we
the enhancement
of
to activation
of the
did
not
observe
any
cytotoxic
activity
when
non-T
cells from
B-CLL
patients
were incubated
with IL 2 sup (data
not shown).
To investigate
whether
could
still
from
the
the
T cell
produce
observed
be present
line,
effects
in the
MLA-144,
IL 2 constitutively.
tive experiment
in the cytotoxic
similar
to that
were
IL 2 sup,
due
we used
which
have
As can be seen
(Fig 4),
activity
to
PHA
that
to
in one representa-
C
Finally,
and
to further
cytotoxic
activity
in T cells
patients.
We investigated
the target
cell specificity
from B-CLL
patients
after incubation
with
be seen in Fig 5, short-term
lytic activity
against
both
target
sup
incubation
NK-sensitive
with
from
Fig
of the
IL 2 sup.
B-CLL
T cells
As can
IL 2 sup induced
and NK-resistant
previous
NK-like
results
demonstrated
activity
in the
that
T cells
B-CLL
patients,
we studied
whether
this cytotoxic-inducer
effect could be explained
by an increase
in the percentage
lymphocytes
with the classic
NK cell phenotype.
In
experiments
not modify
present
in
IL 2
from
of
five
we observed
that incubation
of the IL 2 sup did
the low percentage
of Leu
I 1 cells
(<1%)
the
T-enriched
populations
from
the
B-CLL
patients.
I IILLS
f’JOJBATED
IL-2 SI..P FROM
x,%
%
11%
5.
Cytotoxic
This paper,
ing the nature
10
T
cells
from
81
10
00
of
B-CLL
against
above all, poses an intriguing
question
regardof the IL 2 sup-induced
cytotoxicity
generated
in the T cells obtained
origin
of these
effector
from untreated
B-CLL
cells may be explained
patients.
The
by (a) the
activation
of already
mature
NK cytotoxic
cells;
(b) the
of pre-killer
NK cells into effective
cytotoxic
cells;
or (c) the recruitment
of a different
cell population
not
related
to the NK pathway.
Our data suggest
that this last
differentiation
possibility
is the
most
likely.
this contention.
First, there
basal
levels
of NK activity
The
following
reasons
was no correlation
and the cytotoxicity
support
between
the
acquired
after
incubation
with IL 2 sup. Second,
mature
NK cells
responded
to IFN with an increment
in their lytic activity,
using T cells
enhancement.
from B-CLL
patients,
(Other
authors
have
observation.4’5)
Third,
sup
against
was
active
target
cells.
Finally,
effector
lymphocytes
after the incubation
the cytotoxicity
both
we did not observe
also reported
this
generated
NK-sensitive
there
was no
with
phenotype
with IL 2 sup.
recent
reports
in the
possibility
that, in some systems,
by IL 2 are not typical
mature
that
acquire
cells.’5
LEWS
1;
acquire
LYTIC
4.
10
by the
and
IL 2
NK-resistant
I
I
1
10%
Fig
activity
50
LYSIS
DISCUSSION
Furthermore,
se.
40
SPEOAC
different
target
cells. T cells were incubated
with either complete
medium
(EJ. closed bars) or 20% IL 2 sup (EJ. open bars). After 18
hours
of treatment.
the cultures
were
analyzed
for cytotoxic
activity
against
Chang.
MT-i . Alab. Molt-TG.
Raji. K-562.
Molt-4.
and Jurkat
target
cells.
The data are given
in percentage
of
specific lysis at an effector-target
cell ratio of 50:1.
but
this
WITH
*.A-44
siam
30
%
delineate
cells.
Finally,
since the
induced
strong
s
MLA-144
sup induced
an increase
of T cells from
B-CLL
patients
of IL 2 sup.
strong
MOIT-4
.IURKAT
shown
the molecules
responsible
for the enhancement
of the cytotoxic activity,
we used a semipurified
preparation
of IL 2
obtained
by gel filtration
chromatography
(1 5,000 to 20,000
mol wt). As can be seen in Fig 4, this preparation
also
induced
RAJI
K-%2
supernatants
been
TO
t,OT-
Cytotoxic
activity
UNITS
(LU
of T cells
‘
OELLS)
from
B-CLL
patients
after
treatment
with different
IL 2 preparations.
T cells were incubated
either
with complete
medium
(basal levels) or with the indicated
concentrations
of IL 2 from the T cell line MLA-i44
(a). from a
semipurifled
preparation
obtained
by gel chromatography
(mol wt
1 6.000
to 20.000
daltons)
(b). or from a standard
preparation
(c).
After
1 8 hours
of treatment.
the cultures
were
analyzed
for
cytotoxic
activity
against
K-562 target
cells. The data are given in
lytic
units. LU i O#{149}
cells. as described
in the text.
human
thymocytes
cultivated
de novo cytotoxic
Grimm
et al’#{176}
showed
strong
cytotoxic
mature
NK
cells.
The
literature
support
the
the effector
cells generated
NK cells. We have observed
in the presence
of IL 2 sup
activity
against
tumor
target
that PBMs
incubated
with IL 2
activity
cells. Although
the mechanisms
tions
remain
to be determined,
models
have several
similarities
this report.
For example,
these
by IL 2 in populations
devoid
have neither
the specificity
induction
of cytotoxic
of classic
NK cells
against
NK-resistant
target
underlying
these
observathe effector
cells in these
to those described
by us in
effector
cells can be generated
ofspontaneous
NK
nor the phenotypic
relationship
classic
NK cells is unknown.
Several
pathological
conditions
of these
in addition
effector
activity
markers
cells
to B-CLL
and
of
with
have
From www.bloodjournal.org by guest on January 26, 2015. For personal use only.
ALVAREZ
232
been
found
to
have
similar
immunological
including
an inverted
14/18
mitogen,
and a loss of NK
immunodeficiency
syndrome,
shown
that
the
depressed
abnormalities,
ratio, a poor response
activity.#{176}23 In the
Rook
et a123 have
NK
activity
present
these patients
cannot
be restored
by ‘y-IFN,
to normal
levels by treatment
with IL 2 sup.
identical
to those
it is possible
and
other
with
obtained
that
by us with
the defective
diseases
could
NK
in PBMs
those
B-CLL
patients.
present
Thus,
treatment
of IL 2; (b)
the
in B-CLL;
and
of lymphokines
signifi(c) the
in this
ACKNOWLEDGMENT
thank
studies
are needed
in order to understand
between
the effector
cells mediating
NK
presence
We express our thanks to C. Lorences and 0. Peraile for their
excellent
technical
collaboration,
to Dr Millan for the statistical
analysis, and to Dr J.H. Kehrl for reviewing
the manuscript.
We also
in B-CLL
by in vitro
in the
ET AL
of
IL 2.
Further
relationship
generated
cance
of the T cell abnormalities
potential
immunotherapeutic
use
group of diseases.
but can be raised
These results
are
activity
be overcome
and
to T cell
acquired
recently
DE MON
(a) the
activity
C. Aranda
for technical
assistance
in the
analysis,
Coulter CientIfica,
S.A. (Madrid)
and Dr A. de Ia Hera for his advice.
immunofluorescence
for EPICS-C
facilities,
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From www.bloodjournal.org by guest on January 26, 2015. For personal use only.
1986 67: 228-232
Lymphokine induction of NK-like cytotoxicity in T cells from B-CLL
M Alvarez de Mon, J Casas, R Laguna, ML Toribio, MO de Landazuri and A Durantez
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