PJ, Van Eeden G, Ralph. P. Altrock. BW, Falkenburg .... activation with phytohemagglutinin. J Immunol. 141:3868,. 1988. 35. Metcalf. D,. Nicola. NA: Proliferative.
From bloodjournal.hematologylibrary.org by guest on June 9, 2013. For personal use only.
1989 74: 2398-2404
Interleukin-1 synergizes with granulocyte-macrophage colony-stimulating factor on granulocytic colony formation by intermediate production of granulocyte colony-stimulating factor MR Schaafsma, JH Falkenburg, N Duinkerken, J Van Damme, BW Altrock, R Willemze and WE Fibbe
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InterleukinFactor
I Synergizes With Granulocyte-Macrophage Colony-Stimulating on Granulocytic Colony Formation by Intermediate Production Granulocyte Colony-Stimulating Factor
M. Ronald
By
Schaafsma,
J.H.
Frederik
Falkenburg,
Roel lnterleukin-1
(Il-i
) was
found
granulocyte-macrophage CSF)
on granulocytic
marrow
cells.
cells
colony
depleted
lymphocytes.
Using
we
demonstrated
effect
of
intermediate
cells an
ll-6
did
Furthermore.
this factor to
not
by itself
I
NTERLEUKIN-1 (IL-i) regulation of hematopoiesis. phase response and mediates neutrophils,
clear
phagocytes,3
bone marrow stimulating
factor
and
the
T
was
cells.
not
a
but
by
an
residual experi-
that
observed
Il-i
-
synergy.
a direct
forms, IL-i endothelial
cells’3’4 (G-CSF),
to produce macrophage
CSF- I ), and granulocyte-macrophage whereas T lymphocytes produce
cells,8’2
CSF
to
IL-l.’5
Another nized
hematopoietic
in mice,
proliferation
activity
of IL-i
has been
consisting of a synergy with of early hematopoietic progenitor
which could not yet respond to CSF lL-l was first described in combination
alone.
CSF cells
This
with
bone marrow
cells.20’2’
GM-CSF act cytic/monocytic
synergistically colonies
It was
bone marrow
adherent
IL-i
of but
with IL-3,’7 synergistic IL-i
low
as 0.1
but
not
tion
of
IL-i
Leary
of action. displayed
human
blast
In this
does
tion
From
the
Laboratory
Hematology,
lands;
ofExperimental
University
Laboratory
Hematology.
Medical
cells,
Research,
University
Thousand
Oaks,
of Leuven,
May
Supported
in part
Radiopathology
Normal
Department The
Nether-
marrow
Rega
Institute
for
Medical
hernia
Belgium;
and
cells
Amgen,
/6, and
J. V.D. is research
/989;
accepted
by grants Radiation
July
25,
the
J.A.
from
of the Belgian
Cohen
Institute
for
National
ment
of
requests
Hematology,
to M. Ronald
Bldg
1, C 2-R.
Schaafsma, University
The Neiherland. were defrayed
charge
therefore
payment.
“advertisement” indicate
This
article
in accordance
must with
this fact.
/989 by Grune & Stratton, Inc. 0006-4971/89/7407-OO/9$3.OO/O ©
2398
18 U.S.C.
MD. Medical
Rijnsburgerweg /0, 2333 AA Leiden. The publication costs ofthis article
be section
bone
of IL- 1 was caused effect
between
evidence to synergy cells
IL-i
of
normal phagothat this between
in response
to
our results indicate that proliferation and differentiamyeloid
progenitor
cells.
human
bone
nuclei
AND
After
marrow.
by aspiration
pulposi
patients
collected
in
from
who
informed
consent,
the posterior
iliac
underwent
balanced
Hanks’
heparin
METHODS
(HBSS).
laminectomy.
salt
After
bone crest
solution
dilution
of The
containing
in
RPMI-1640
5% fetal bovine serum (FBS; Hyclone, UT), the cell suspension incubated with 125 mg sterile carbonyl iron powder (Grade SF; Aristopharm, DeIft, The Netherlands) in Erlenmeyer glass flasks was
of reprint
IL-i (CD34indirect
CD34-enriched
accessory
Furthermore, the in vitro
was obtained were
(45
Research
Fund. Address
between
with
/989.
Protection.
associate
from
lineage-specific
preservative-free
CA.
Submitted
as
unclear a direct
mono-
Leiden.
Leuven,
the
as well
colony growth depleted of mononuclear
by residual
GM-CSF. not affect
in
it was shown that IL-6, but not support of IL-3--dependent
We present and attributed
is indirect,
of committed,
Center.
of Immunobiology.
of synergy
the synergistic
MATERIALS
of
M-CSF
of highly purified cells, indicating an
in
granulocytic
produced and
proliferacells
an indirect effect due to an factor produced by accessory a lack
lymphocytes.
Il-i.
cells.
with
formation
marrow T
of IL-i
IL-I IL-6
or
we show
as
that
progenitor
in combination
on
bone
G-CSF
indicate
suggesting that the synergy production of IL-6.
report
synergistic of
From these studies it remained between IL-I and CSF was
cell colony
and
this
results
pro-
GM-CSF.
Inc.
Furthermore, synergy
GM-CSF
human
GM-CSF
of accessory
on HPC, intermediate
marrow cells, by intermediate
and
G-CSF with
GM-CSF-dependent
on colony growth human progenitor
mode IL-l,
colony
-induced
synergy
myeloid
et a122 reported
IL-3 enriched) and
synergisti-
an anti-G-CSF
concentrations
& Stratton.
or IL-3.2’ the synergy of
the
numbers
growth
colony
with Il-i
Our
mature
it act monocytic
the
at
U/mI).
stimulates
by Grune
G-CSF,
and
(10
of small
GM-CSF whether
and
detected
ng/ml
did or
that for
of G-CSF
be
11-6,
1989
cytic
showed
relatively
activity
of granule. human non-
enhances
could
cytes
on human
that
proliferation of normal
cells,#{176} and that
on the (HPC),
M-CSF,’6
demonstrated
on the in cultures
recog-
synergy
analogous activities have now been documented GM-CSF,’8 and G-CSF.’9 Recently, a similar effect between IL-I and CSF has been reported
combinations
activity
cells.
and
(GM-CSF), in response
GM-CSF
Using
nor
granulocytic
responsible
IL-I-induced
granulocyte colonyCSF (M-CSF or
W. Altrock.
experiments
antibody was
action
during mononu-
Bruce
growth.
on
Neutralization
monoclonal
0
Damme,
colony
GM-CSF
presence
stimulatory
observed induces
on CFU-GM with
duction
has several activities in the IL-l is involved in the acute the increase in circulating
fibroblasts,7
stromal
formation.
showed
neither
immature Furthermore,
including
infections.’
bone
. Neutralization
to
had
human
produced
Il-i
contribute
effect cally
marrow
Jo Van
E. Fibbe
with
of Il-i
antiserum
Duinkerken,
Willem
(GM-
bone
activity
and
factor phagocytes
progenitor
anti-ll-6
11-6
microbial
of normal
hematopoietic in response
using
induced
growth
that
on
ments
synergistically
of mononuclear
action
accessory
act
CD34/HIA-DR-enriched
direct
an
to
colony-stimulating
Nelleke
Willemze.
of
minutes; the
Depart-
mononuclear
Center,
suspension myeloid
in part by page hereby 1734
solely
to
particles
were
removed i,000
harvested serum
rosetted
with
and (FBS).
magnet The
Blood,
was
added
cells were iron-loaded
by sedimentation
through
g; 20 minutes,
20#{176}C),and
washed
times
three
T lymphocytes and sedimentation
Vol 74. No 7 (November
in
were
RPM!
the
the cell
collected,
and
mononuclear interphase plus
removed
5%
cells fetal
by being
(AET)-pretreated
of the rosetted
15),
remove and
Ficoll-Isopaque
the
then
bromide
phagocytosis
To
cells, and residual
2-aminoethylthiouronium
erythrocytes
to allow
phagocytes.
for 5 minutes.
red blood
were
g/cm3,
agitation)
mononuclear a sterile
was shaken cells,
bovine sheep
continuous
by
phagocytes,
phagocytes (I .077
marked
37#{176}C, with
iron
1989:
cells through
pp 2398-2404
From bloodjournal.hematologylibrary.org by guest on June 9, 2013. For personal use only.
IL-i
SYNERGIZES
WITH
Ficoll-lsopaque. than
The
GM-CSF
nonrosetting
1% T lymphocytes using
cence goat
anti-mouse
The
anti-CD3
(Becton
Immunological
7TD1
2% mononu-
a-naphthylbutyrate
esterase
bone
cells was obtained
forming
cells using
an anti-CD34
Mountain
View, CA),
and
a fluorescence
Dickinson). and
T lymphocytes, at 4#{176}C. The
labeled
with
and
incubated
were
incubated
MoAb
Flow Laboratories,
separation
was
performed
negative
and
cence
intensity second
50%
centrifuged
FBS,
only.
The
(150
g;
40%
(or
and
stimulated
U/mL),
were
were
0.5
were
l0
of greater
than
Normal
defined
iO bone
Reagents. provided
MA).
Recombinant
Amgen
from
purification
human
crude
other
cytokine
CSF,
and
human
as low
neutralized
by
pure
magglutinin
(PHA).24
NH2-terminal
amino
IL-6
against was
The acid
purity sequence
of
to detect
in
protein
only
(Fig
with 1).
numbers of granulocytic colonies numbers of monocytic colonies were
cultures
IL-I-stimulated
and controls
This
attributed
in the
the
of
of both
as compared
be completely
(Fig simi-
(Fig 2B).
-
100
-
I.-
IL-i
0 ‘4-
0 50
-
was
contain
control as
levels
sheep
of
IL-lfl
could Standards IL-l$.
supernatants with wa(
Recombinant
of
phytoheverified human
010
0 be
(kindly
recombinant from
the
0
from
of the
activity,
stimulated
analysis.
was
endotoxins,
for Biological
cells
U/mL),
could
numbers
presence
GM-CSF
interferons,
activity raised
obtained
mononuclear
0
IL-ifl not
IL-2,
able
human
higher
in the
100 with
.05)
in the ±
was obtained
biologic
(P
numbers
C
It did
assay
The
in an assay
by Dr S. Poole,
peripheral
lysate
and
a
.c
0 0 0
U/mg)
Institute,
IL-ia,
to be free
amoebocyte
(mean
preparations
including
aggregates
(l0
previously.4
as 60 pg/mL.
Electrophoretically human
found
as measured
and Control,
lymphokine
increase
were
obtained
stimulated
in colony
whereas for
(10
determine
of recombinant and in the presence
were
GM-CSF
cultures
To
IL-i and GM-CSF, nordepleted of mononuclear
0)
I0
plated.
U/mg)
Wil-
150-
microscope.
146
Electrophoretically
tested,
It was
by a Limulus
endotoxin preparation, completely
(l0
as reported
activities
!L-6.
(Genetics
G-CSF CA).
cells
the
using
0
of
colonies
GM-CSF
(rh)
Clark
each
number
an inverted
10 CFU-GM
mononuclear human
like
100%
cells, the
±
of !L-6. tested,
factors
or monocytic
are 503
evaluated
GM-CSF.
between cells,
Significantly
colonies
200
T lymphocyte-
days,
using
U/mL
marrow
Oaks,
10
for day
100
schedule
provided
scored
by Dr S.C.
(Thousand
obtained
were
Recombinant
kindly
After
to an
lar
into the wells of microtiter
as granulocytic
in our laboratory
of GM-CSF
23) per
well.
U/mL).
difference
or
mL,
CD34/HLA-DR-enriched
each
20 cells,
values
presence
tested
in
U/mL
(100
the
of 0.1 and
cytokines
were
T lymphocytes,
FBS,
lL-ij3,
no stimulatory
phagocyte-
(10
2A),
of GM-CSF
replicates
cells were plated
containing
colonies,
Six
2,000
other
in a
At a dilution
in a-MEM.
determining
control,
medium.
mononuclear
marrow
G-CSF,
concentration
in
20%
(2.25%)
controls,
As a negative
plated
CFU-GM
-
x
In cultures
plates.
n
level.
bone
GM-CSF,
a plateau
plus
humidified
containing
bone and
CFU-GM IL-l and control
culture.
in a fully
solution
as positive
to the culture
containing
cells
with
included
growth added
depleted
containing
F!TC-
in RPM!
in CFU-GM
cultured
of a methylcellulose
Cultures colony
a-MEM
40%
the
20#{176}C),resuspended
plated
of 37#{176}C and 5% CO2 in a medium
a-MEM
IL-6),
between
collected
10 minutes,
a-MEM 5% FBS, and immediately CFU-GM culture. Cells were atmosphere
were
was raised
IL-6.2’
with
and
IL-I
effect marrow
of increasing concentrations CSF (0, 1, 10, 100 U/mL), IL-i
on the fluoresthe
human
neutralize
Differences
between
phagocytes
medium
with
of rhG-CSF
pairs.
synergistic
human
mal
times,
in a CD34
point
based
of
was able
and CSF.
analysis.
Synergy
an anti-
separated
antiserum
pure
cross-react
interferons,
activity
anti-IL-6
to completely not
test for matched
possible
with 5% FBS. Cell argon-ion laser beam
cut-off
IL-l,
proliferation
Amgen)
(iSA,
RESULTS
times.
two
essential
incubated
Cells
three
washed
were
able did
!L-6-dependent
It did not show cross-reactivity
of 1:200. The
of The
30
immunoglobulin washed
was chosen, sample
for
it was
the
the colony-stimulating
5% FBS,
at 4#{176}C with
FACS, cells
fraction.
antibody
MoAb in RPM!
minimal
the
fraction
a control
conjugated
times
MoAb
electrophoretically
antiserum
coxon
using half-maximal
or rhIL-3.
against
Statistical
Becton phagocytes
anti-CD34
Scotland)
positive
negative
of
FACStar,
anti-mouse
Irvine, using
the positive
IL-la,
two
Eagle’s
HLA-DR
and a CD34
positive
Dickinson)
30 minutes
in a-modified
nm.
(Becton
to phycoerythrin,
(a-MEM;
at 488
MoAb
at 4#{176}C, and
for
conjugated
resuspended
The
(FACS;
goat
for 30 minutes
rhGM-CSF
Dickinson,
with
FITC-conjugated
cells
tuned
at a dilution
rabbit
quantitated 1 U giving
anti-G-CSF
neutralize
U/ml)
of 1:300
of progenitor
was
7TD1?’
( I ,000
Becton
of mononuclear
cells were washed
(Nordic)
HLA-DR
sorter
cells, depleted
were
minutes
Next,
cell
selection
for
line
to completely
My-lO;
(HPCA,
an anti-HLA-DR
activated
Enrichment
cells.
by positive
MoAb
Bone marrow
antiserum
marrow
IL-6
cell
Aarden (Central Laboratory Transfusion Service, Amsterdam,
Blood
cells. The murine
with
CD34/HLA-DR-enriched
of Dr L.A.
gift
Red Cross
Netherlands).
Dick-
staining. colony
was a kind
murine
and less than
with
IL-6-’
Netherlands
(FITC)-labeled
(Nordic
Netherlands),
less
immunofluores-
isothiocyanate antisera
as determined
contained
indirect
(MoAb)
and fluorescein
Tilburg,
phagocytes
cells
with
immunoglobulin
Laboratories, clear
antibody
CA)
2399
INDUCTION
interphase
as determined
monoclonal
inson, Oxnard,
BY G-CSF
by
GM-CSF
100
(U/mi)
Fig 1 . Synergistic effect of Il-i (10 U/mI) on GM-CSFsupported CFU-GM colony formation of normal human bone marrow cells. depleted of mononuclear phagocytes and T lymphocytes. Data are a percentage (mean ± SD of 1 0 experiments) of control CFU-GM growth obtained in the presence of GM-CSF 100 U/mL.
From bloodjournal.hematologylibrary.org by guest on June 9, 2013. For personal use only.
SCHAAFSMA
2400
200
ET AL
200
-
.C
0
150
150-
I-
Fig2. EffectoflL-1 (10 UI mL) on (A) GM-CSF-supported CFU-G and (B) cFu-M colony formation of normal human bone marrow cells. depleted of
>1
C
0 0 100
U
100
IL-i
0
mononuclear
0 0 50
50
0
control 0
00
1
A
GM-CSF
done,
by residual using
purpose
of highly
suspensions
depleted
bone
of accessory
additional
enriched
for
HPC.
and
T
were
further
cell suspensions
and
studies
purified
phagocyte-
marrow cells
cells,
were
For
this
depleted cells,
from
cultures
GM-CSF
tion was no longer was
present
using
cells,
(Fig
the synergy
colony
3), suggesting
that
forma-
the effect
indirect. Characterization
br.
In
order
factor
responsible
CSF,
neutralization
duction
of to
of IL-6
known
hematopoietic
centrations IL-i
progenitor on granulocytic
and
the
further for
the
synergistic
characterize synergy
experiments
is induced
the between
were
IL-i-induced and GM-
performed.
we added
antiserum
Control
Since
prohas con-
IL-6
neutralizing
to cultures
measurements
.c
fac-
IL-l
by IL-i#{176} and since
activities,
of an anti-IL-6 GM-CSF.
IL-i-induced
colony
with no
effect
colony
IL-i
in the presence
of the
antiserum
by IL- 1 was found
growth
was
Control
preparations
calculated Since fined
the
not
IL-i effect
colonies, the
by
further
GM-CSF
of
Further-
addition
of
synergistically
Results
were
for rh-IL-6
(data
proved
active
and
and
GM-CSF
we considered
that
the
contained was
G-CSF
addition MoAb,
the conas the
of neutralizthe synergy
and GM-CSF was totally abrogated (Fig 5). was not due to toxicity of the antibody since
GM-CSF-stimulated To
and
effect. Following of an anti-G-CSF
not
IL-6 at concentrano direct effect on
of IL-6
biologically
data
(Fig 4A).
Also, had
IL-6
measurements
to granulocytic
between
purified
were
could
colony verify
that
growth the
be mediated
was
not diminished.
synergy
between
by IL-i-induced
150#{149}
150
-
125#{149}
125
-
IL-i
and
production
of
0
100
100-
IL-i
>‘
C 0
Fig 3. Effect of IL-i (10 UI ml) on (A) GM-CSF-supported CFU-G colony formation of normal human bone marrow cells. depleted of mononuclear ph.gocytes and T lymphocytes and on (B) CD34/HLA-DR-onriched bone marrow cells. Data are a percentage (mean ± SD of two experiments) of control CFU-G growth obtained in the presence of GM-CSF 100 U/mI.
0
75.
U
75
-
IL-i
0
control
C 0 U
50.
50
-
25
-
control
0
25#{149}
0
00
A
1
GM-CSF
10
(U/mI)
T
of anti-IL-6
enhancement
influenced
amount of the cytokine. synergy between IL-l
factor mediating ing concentrations This
human
and
U/mL,
on the
growth
for both
shown).
IL-6
containing
in supernatants
(
C
antHL-6
:::
.2 0
IL-i
C,
.
antHL-6
100
50
0 0110
A
100
GM-CSF
(U/mI)
GM-CSF
(U/mI)
200
-
control
IL-i
150
a::::
IL-6
IL-i
#{149} IL-6
100 Fig 4. Effect of neutralization of the Il-iinduced 11-6 production (A). and effect of addition of exogenous natural 11-6 (30 U/mL) (B) on the synergy between IL-I (10 U/mI) and GM-CSF on CFU-G colony growth of normal human bone marrow cells. depleted of mononuclear phagocytes and T lymphocytes. Data are a percentage (mean ± SD of (Al two and [BI six experiments) of control CFU-G growth obtained in the presence of GM-CSF 100 U/mL
50
0
100
0JI,
B
I 75 .c
150
-
control
0
IL-i
a-
125 >a
C
0 0
L#{149}anti-G-CSF IL-i
100
#{149} anti-G-CSF
0
I
______ _
25
0
i
GM-CSF
10
(U/mi)
100
Fig 5. Effect of addition of a neutralizing anti-G-CSF MoAb on the synergy between IL-i (10 U/mi) and GM-CSF on CFU-G colony growth of normal human bone marrow cells. depleted of mononuclear phagocytes and T lymphocytes. Data are a percentage (mean ± SD of four experiments) of control CFU-G growth
obtained
of GM-CSF
in the
100 U/mI.
presence
From bloodjournal.hematologylibrary.org by guest on June 9, 2013. For personal use only.
2402
SCHAAFSMA
Table
1 . Synergy
Between
G-C SF and
GM-CSF
on Hums n CFU-G
Colony
ET AL
Growth
GM-cSF Me6um
1 U/mL
Medium
0±1
6 ± 10
G-CSF1U/mL
2±3
i3±i9
70±23
103±10
G-CSF1OU/mL
2±3
16±26
97±31
125±33
G-CSF100U/mL
27±31
G-CSF
1,000U/mL
(mean
Data are a percentage
cells
marrow
G-CSF,
depleted
were
additional
binations
±
in
CFU-G
I
present
were
performed
G-CSF and synergizes
G-CSF
growth.
concentrations
This
of G-CSF
effect
as low
as
cell suspensions
already
140
253 owth
1 had
GM-CSF
was
illuson
exogenous
IL-6,
and
was
marrow
no effect
at still
cells,
(Table
on
2).
U/mL,
study
GM-CSF
clear
from
bone
lL-l
and
colonies
and
and
enriched for CD34/HLA-DR
HPC,
cell.
mediated
by
Only
When less
anymore.
bone
2%
results
by IL-l.
IL-6
since or
was
synergizes
with
only
of IL-3
gistic
effect
enhance
GM-CSF-stimulated
colony formation. When was neutralized by a rabbit CFU-GM
of
IL-l-induced IL-6 anti-IL-6 antiserum,
in the
colonies
presence
prothe
of IL-i
and
GM-CSF this
between
both
Synergy Colony
Between
G-CSF
Growth
and GM-CSF
on Human
CFU-G
of CD34/HLA-DR-Enriched Marrow
Bone
enhanced
either
factor
Medium
1 U/mI
10 U/mI
100 U/mI
that IL-3
0
0
72
100
0
0
70
107
0
0
109
148
murine plexity
10 U/mL
G-CSF
iOOU/mL
G-CSF
i.000
U/mL
Data are a percentage presence
of GM-CSF
70
237
298
differential
430
452
maturity
of control
CFU-G (results
colony
growth
of one experiment).
obtained
in the
IL-l and GM-CSF formation, we considIndeed, antibody
in
that
attributed
to a synergy
the
neutralno syner-
increase
in
between
In accordance
with
and
growth
to support
cells,
suggests
a
with
may
colony
direct
effect
myeloid of the
activities
by different
IL-3
synergy
recent
M-CSF
formation.
that with
both IL-3
marrow,
on proliferation
progenitor regulation
The
pres-
with
G-CSF” on blast and
and cell
Leary
These factors
of early
by Bartelmez
a direct the
bone
et
studies that act HPC.
In
that G-CSF, but not IL-6, GM-CSF on relatively mature
studies
exert on
activity
with
on HPC.
bone
we demonstrate
also
signifi-
in comparison
CD34/HLA-DR-enriched
of murine
study
previous
GM-CSF
effect was already observed at low (10 U/mL), which, in the absence on
in the
and
348
the
to the cells. It in the
a similar effect of IL-6 in humans. that IL-6 and probably G-CSF are
IL-i
43
whether
T lymof the
colony
synergy
In addition,
276
100 U/mL
failed
synergistically
IL-i G-CSF
CFU-G
development
HPC.
clear
is a co-stimulant
In mice, Ikebuchi et al described IL-63’ have a synergistic activity
Medium 1OU/mL
IL-6
of G-CSF
this
of
is involved GM-cSF
This
T cells, and activated for the production between colony
was tested.
alone. This of G-CSF
of GM-CSF,
the present
Cells
CSFs
cantly
colony
cells.
indicating that on CFU-GM
indicating
reports,3#{176} the combination
ence
colony and
and IL-l-induced G-CSF. Further evidence for obtained in additional studies, in which synergy
was
suggest 2.
was
IL-6
be attributed of accessory
this activity. an anti-G-CSF
found,
colonies
a122 found Table
was
granulocytic
marrow
not
that
to mediate experiments with
ization
the
it is not
synergistic activity. the synergistic effect seen on granulocytic
indicate
did
recently
ered G-CSF
and IL-6
production mononuclear
present
of
30 to i ,000
progenitor
recent studies,333 with GM-CSF
of PHA-stimulated may be responsible
Since
that
indirect factor.
of myeloid
At
from
GM-CSF-stimulated synergy between
no
reported
concentrations
number
Bone
addition
ranging
in
formation of early blast cell colonies?1 and a similar synergistic effect with GM-CSF has been reported on myeloid (CFU-GM) colony growth.3U3 However, our results CFU-GM duction
U/mL.
100
Furthermore,
has been phocytes
mononuclear were further
suggest
GM-CSF was of an intermediate
diminished.
in concentrations
presence
observed
on the
IL-6
GM-CSF
of
activity of IL-6 could or absence of small numbers
of
marrow
not
formation.
of
antibodies against the sorting, no synergistic
that we considered, endothelial cells?
phagocytes#{176} is induced
formation
these
than
These and
production
was one of the factors IL-6 by fibroblasts,28
that
± 185
302
presence
on proliferation
activation
of mononu-
by the combination effect on a committed,
monoclonal and cell
IL-i
the
depleted
1% T lymphocytes,
using antigens
between
and GM-CSF act human CFU-GM
lymphocytes.
less than
found
was
synergy
T
containing
suspensions,
effect
suspensions
progenitor
phagocytes
IL-l of
was enhanced suggesting an
GM-CSF,
lineage-specific cell
marrow
phagocytes
granulocytic
in the
finding is in contrast with IL-6 may act synergistically differences
we demonstrate that on the proliferation
synergistically
colonies
184±47
i76
±
did not influence Thus, we found
growth.
colony DISCUSSION
In this
obtained
com-
detectable
10 U/mL bone
of IL-
using
GM-CSF. As with GM-CSF
was
on CD34/HLA-DR-enriched
which
±
100
145±34
SD of three experiments) of control CFU-G colony phagocytes and T lymphocytes.
experiments
Table
colony
198
100 U/mI
10
±
of mononuclear
of recombinant
trated
45±23
± 102
161
10 U/mI. 69
et a142 suggest
synergistic
proliferation
of
activity very
with
primitive
cells. This illustrates the comof hematopoiesis, suggesting a
of various
cytokines
depending
on
level of the target cell. Thus, the indirect synergistic of IL-I on primitive and mature HPC are mediated intermediate
factors.
the
From bloodjournal.hematologylibrary.org by guest on June 9, 2013. For personal use only.
IL-i
SYNERGIZES
WITH
GM-CSF
BY G-CSF
2403
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