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1994 84: 608-615
Interleukin-4 receptor regulation in human monocytic cells
H de Wit, DW Hendriks, MR Halie and E Vellenga
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Copyright 2011 by The American Society of Hematology; all rights reserved.
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Interleukin-4 Receptor Regulation in Human Monocytic Cells
By Harry de Wit, Dick W. Hendriks, M. Ruud Halie, and Edo Vellenga
The regulation of
the interleukin-4 receptor (IL-4R) was studied at mRNA andprotein level in
monocytic cells on stimulation with activators of dfierent intracellular signaling pathwaysandIL-4.ActivationofproteinkinaseC-dependent
pathways with phorbol myristate acetate (PMA) or activation of protein kinase A-dependent pathways
with DBcAMP
and prostaglandinEz resulted in an augmented IL-4R expression at mRNA and protein level. Transcriptional and posttranscriptional mechanisms seemed to be involved in the
promotive effect of DBcAMP because the transcription rate
increased 1.8-fold, andthe half-life of IL-4R mRNA was prolonged to 150 minutes compared with 120 minutes in unstimulated cells. In contrast, the effect of PMA could only
beascribed to changes at transcriptionallevel.However,
activation of Ca2+-dependent
pathwayswith A23187 or stimulation with IL-4 had no effect onthe IL-4R expression. The
unresponsivenessto IL-4 couldnot be ascribed to a nonfunctional receptor because IL-4 did modulate the CDl4, CD23,
and HLA-DR antigen expression. These results are
in contrast with IL-4R regulation in T cells, which is affected by IL4- and Caz+-dependent pathways. The discrepancy might
be caused by the presence of the common IL-2 receptor y
chain (yoye)in T cells and the absence of the yc in monocytic
cells,ashasbeenshownbypolymerasechainreaction.
These data indicate that IL-4Rs are differentially regulated,
depending on the cell type studied.
0 7994 by The American Societyof Hematology.
I
ing pathways affect the IL4R expression in this cell type.
For these studies, we used the Mono Mac 6 cell line because
these cells havemany properties in common with normal
human monocytes.” The results show that protein kinase A
( P a ) - and PKC-dependent pathways are involved in the
regulation of the IL-4R gene expression, whereas Ca” mobilization and stimulation with IL-4 had no effect.
NTERLEUKIN-4 (IL-4) a product of activated T lymphocytes affects, among others, B cells, T cells, mast cells,
and hematopoietic progenitor
The divergent activities of IL-4 on these cell types are mediated through binding
of IL-4 to a high-affinity receptor. Receptor studies with Iz5Ilabeled IL-4 on resting lymphocytes initially identified a
trimolecular complex consisting of a 65/70-kD doublet and
a 120-kD protein with high-affinity binding sites (Kd, 100
pmol/L).6-8More recently, it has been shown that the polypeptide chain with a molecular weight of 70 kD (p70) was
the breakdown product of ~ 1 2 0In. ~addition, recent investigations have indicated that interleukin-4 receptors (IL-4Rs)
share a common y chain ( y c ) with the IL-2R and the IL7R.10-12
cDNA encoding the murine and human IL-4Rs have been
isolated, characterized, and seemedto be member of the
recently described superfamily of cytokine receptors because
of the conservation of a 4-cystein motif in the N-terminal
region.’”’‘ Studies onIL-4R regulation inhuman T cells
have shown that IL4R expression is controlled by different
intracellular signaling pathways. T cells stimulated with the
protein kinase C (PKC) activator, phorbol myristate acetate
(PMA), or the calcium ionophore, A23187, showed an increased expression of IL4R at mRNA and protein level.’’
In addition, IL-4-stimulated T cells showed an increased
IL4R mRNA expression by a factor 10, which wasindependent of PKC or CaZ+m o b i l i z a t i ~ n . ’ ~ ~ ~ ~
In view of the profound effects of IL-4 on monocytic
functions, we questioned whether similar intracellular signal-
From the Division of Hematology, Department of Internal Medicine, University of Groningen. Groningen. the Netherlands.
Submitted October 14, 1993; accepted March 18, 1994.
E.V. is a fellow of the Royal Netherlands Academy of Arts and
Sciences.
Address reprint requests to Ed0 Vellenga,MD,Department of
Internal Medicine, University Hospital, Oostersingel 59, 9713
E2
Groningen, the Netherlands.
The publication costsof this article were defrayed in part by page
chargepayment. This article must therefore be hereby marked
“advertisement” in accordance with 18 U.S.C. section 1734 solely to
indicate this fact.
0 1994 by The American Society of Hematology.
0006-4971/94/8402-0017$3.00/0
608
MATERIALS AND METHODS
Cell culture and stimulation. Mono Mac 6 cells (gift ofDr
H.W.L. Ziegler-Heitbrock, Heidelberg, Germany)” were cultured in
RPMI-1640 medium (GIBCO-BRL, Gaithersburg, MD) supplemented with15% fetal calf serum (FCS; Hyclone, Logan, Utah),
nonessential amino acids (GIBCO), 1 mmoYL pyruvate (Sigma, St
Louis, MO), 1 mmom oxalacetate (Sigma), 2 mmol/L L-glutamine
(GIBCO), and antibiotics. Eighteen hours before each experiment,
cells were washed and resuspended in fresh medium at a cell concentration of 2 X 1O6/mL.
Peripheral blood cells were obtained from volunteer platelet
donors and mononuclear cell suspensions were prepared by FicollHypaque density-gradient centrifugation. T lymphocytes wereremoved by E-rosetting with 2-aminoethylisothiouronium bromidetreated sheep red blood cells. Monocytes were further enriched by
plastic adherence for 1 hour at37°Cand showed a purity greater
than 95%, detected by fluorescence-activated cell sorter (FACS)
analysis withan anti-CD14 monoclonal antibody (MoAb; Becton
Dickinson, Sunnyvale, CA) and staining. Monocytes were cultured
in RPM1 1640 medium containing 10% FCS and antibiotics.
The following activators were used in the experiments: the Caz+ionophore A23187 (Sigma), PMA (50 nglmL; Sigma), 2”dibutyrylcyclic adenosine monophosphate (DBcAMP;
mom; Boehringer, Mannheim, Germany), prostaglandin Ez (PGE2; IO-’ mol/L;
Boehringer), actinomycin-D (10 pg/mL; Sigma), and rhIL-4 (gift
from Dr P. Trotta, Schering Plough, Bloomfield, NJ) with a specific
activity of lo7 U/mg. The viability of monocytes and Mono Mac 6
cells after stimulation was greater than 95% in all experiments.
mRNA analysis. Total cellular RNA was isolated using the guanidium isothiocyanatekesium chloride method. Twenty-microgram
samples of total cellular RNA were size fractionated on 1.1% agarose
gels with 2.2 m o m formaldehyde and blotted onto nylon membranes
(Hybond N+; Amersham, UK).**
cDNA probes were labeled with [’*P]deoxycytidine triphosphate
(3,000 Cihmol; Amersham) using the hexanucleotide primer technique.” The following cDNA probes were used: the human IL4R (gift from Dr M.K. Spriggs, Immunex, Seattle, WA), human
macrophage colony-stimulating factor (M-CSF) (gift ofDr S.C.
Blood, VOI 84, NO 2 (July 15). 1994: pp 608-615
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IL-4R REGULATION IN MONOCYTIC CELLS
PMA
A
time (h)
609
3 4 5
0
7 9 1 1
IL-4R
28s
DBcAMP
B
time(h1
0
1
3
6 1 2
1L-4R
28s
C
(SSC), 0.1% SDS (65°C. 30 minutes), I X SSC, 0.1 Olr SDS (65°C.
30 minutes), and finally in 0 . 3 SSC,
~
0.1% SDS (65°C. 30 minutes).
Membranes were then exposed to x-ray films (Kodak X-Omat XAR:
Eastman-Kodak, Rochester, NY) at-80°Cusingan
intensifying
screen. Quantification of mRNA levels was performed by densitometry using a gel-scan laser densitometer (Pharmacia LKB, Uppsala,
Sweden).
Run-on transcription assav. The transcription rateofthelL-4R
In short, nucleiof 50
gene was studied as described previo~sly.'~
X IO6 cells were isolated and resuspended in transcription buffer
containing ['*P]-uridine triphosphate (3.OOO Cilmmol; Amersham)
and incubated at 26°C for 20 minutes. Transcription was terminated,
and nuclear RNA was isolated and hybridized to linearized pGEM
and to linearized plasmids containing the cDNA inserts for the IL4R and 28s rRNA. Hybridization. washing, exposure, and quantification of transcription rates was performed as described above.
Reverse transcription (RT)and polvmerase chain reaction (PCR).
A total of 0.5 pg total cellular RNA was incubated with 135 mmoll
L Tris-HCI, pH 8.3; 204 mmol/L KCI; 27 mmol/L MgC12;0.24 mgl
mL bovine serum albumin; 5.4 mmol/L of each deoxynucleotide
triphosphate; 14.3% glycerol; 3 mmol/L dithiothreitol; 0.2 pg
pd(N),; 5 U RNAguard (Pharmacia); and 3 U MoloneyMurine
Leukemia virus reverse transcriptase (Pharmacia) in a total volume
of 15 pL for I hour at 37°C.
Specific primer pairs for the common yc and for theGAPDH
gene were synthesized on a Gene Assembler Plus DNA synthesizer
(Pharmacia) and purified using NAP-l0 columns (Pharrnacia). One
microliter of 50-pmoVL 5' and 3' primers were added to each reverse
transcriptase (RT) reaction sample and DEPC H20 was added to a
final volume of 50 pL. The samples were then heated for 5 minutes
at 95°C and placed on ice; 5 U of Taq DNA polymerase (Pharmacia)
was added, and each sample was overlaid with 50 pL paraffin oil.
The reaction tubes were then placed in a thermal cycler (Pharmacia),
and polymerase chain reaction (PCR) was performed for 20, 25, 30.
c
0
PGE2
U
time(h1
0
A23187 (h1
0 3 6 1 2
6
IL-4R
IL-4R
28 S
M-[SF
Fig 1. Mono Mac 6 cells were stimulated with (A) PMA (50 ngl
mol/L), and (C) PGE2 (lo-' mollL).Atthe
mLI, (8) DBcAMP
indicated time points, total cellular RNA was analyzed for the IL-4R
mRNA expression. Rehybridization with a 28s probe was assessed
to control RNA loading in each lane.
Clark, Genetics Institute. Cambridge, MA).and
the linearized
pBr322 plasmid containing a 7.8-kb human 28s cDNA insert.
Hybridization was performed at 65°C for 18 hours in 0.5 mom
Na2HP04, pH 7.2, 1 mmol/L EDTA,7% sodium dodecyl sulfate
(SDS).Membranes were washed in 2 x saline sodium citrate buffer
28s
Fig 2. The effects of the calcium ionophore A23187 (5 pmollL) on
lL-4R mRNA expression. Mono Mac 6 cells were stimulated at the
indicated time points, and the filter was hybridized with cDNA probes
for lL-4R, M-CSF, and 28s.
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IL-4R REGULATION IN MONOCYTIC CELLS
A
61 1
Control
PMA
OBcAMP
28 S
B
120
Relative rnRNA
* Control
* PLU
100
*
DECAMP
80
60
40
0
0
30
60
90
120
150
180
Time (min)
Fig 5. Experiments to determine the half-life of IL-4R mRNA. (A) Mono Mac 6 cells were stimulated with medium, PMA (50 nglmL). and
DBcAMP
mollL) for 6 hours. Then actinomycin-D(10 pg/mL) was added to inhibit transcription. At various time points after actinomycinD addition IL-4R mRNA expression was detected. (B)Graphic analysis of IL-4R mRNA half-life.
ficity of the DBcAMP effect, cells were stimulated with
PGE2 (
mol/L), which elevates intracellular CAMP levels.*'As depicted in Fig IC, stimulation with PGE2 for 6
hours showed a twofold increase in IL-4R mRNA expression. In contrast, Mono Mac 6 cells stimulated with the
calcium ionophore, A23187 (1 and S pmol/L), showed no
change in IL4R mRNA expression (Fig 2). This could not
be ascribed to inadequate stimulation because rehybridization with an M-CSF probe showed a strong increase in MCSF transcripts. Furthermore, the effects of IL-4 on the IL4R expression were studied. Mono Mac 6 cells did not show
a change in the 1L-4R mRNAexpression on IL-4 stimulation
(Fig 3A). However, the IL-4R was functional because IL4-stimulated cells showed an increased CD14 antigen expression of 40%. whereas the HLA-DR antigen expression
decreased to 30% after IL-4 stimulation. To exclude the
possibility that the unresponsiveness of the IL-4R for IL-4
was related to the used cell line, monocytes were stimulated
with IL-4. As depicted in Fig 3B, no change in IL-4R mRNA
expression was noticed. In three separate experiments,
monocytes showed no significant increase in IL-4R mRNA
expression on stimulation with IL-4 for 6 hours ( I .O I -fold
-t 0.09-fold increase compared with unstimulated monocytes
(mean t SEM; n = 3)).
Anulysis of IL-4R transcriptionrutes.
Run-on assays
were performed to determine whether the effect of PMA and
DBcAMP involves an increase in the transcription rate of
the IL-4R gene. A total of SO X IOh Mono Mac 6 cells were
stimulated with medium, PMA, and DBcAMP for S hours.
Unstimulated Mono Mac 6 cells showed a spontaneous transcription of the IL4R gene, which is in accordance with the
constitutive mRNA expression of the IL4R. In three separate experiments, PMA- and DBcAMP-stimulated cells
showed a 1.33-fold -t 0.13-foldand1.81-fold 5 0.32-fold
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IL-4R REGULATION IN MONOCYTIC CELLS
A
61 1
Control
PMA
DBcAMP
28 S
B
120
Relative m R N A
8 Control
PMA
*
100
* DBeAN?'
80
60
40
2o
0
t
0
180
30150
60
l20
90
Time (min)
Fig 5. Experiments to determine the half-life of IL-4R mRNA. (A) Mono Mac 6 cells were stimulated with medium, PMA (50 ng/mL), and
DBcAMP (lo-' mol/L) for 6 hours. Then actinomycin-D ( l 0 pg/mL) wasadded to inhibit transcription. At various time points after actinomycinD addition IL-4R mRNA expression was detected. (B) Graphic analysis of IL-4R mRNA half-life.
ficity of the DBcAMP effect, cells were stimulated with
PG& (
mol/L), which elevates intracellular CAMP levels.*'As depicted in Fig IC, stimulation with PG& for 6
hours showed a twofold increase in IL4R mRNA expression. In contrast, Mono Mac 6 cells stimulated with the
calcium ionophore, A23187 (1 and 5 pmol/L), showed no
change in IL4R mRNA expression (Fig 2). This could not
be ascribed to inadequate stimulation because rehybridization with an M-CSF probe showed a strong increase in MCSF transcripts. Furthermore, the effects of IL-4 on the IL4R expression were studied. Mono Mac 6 cells did not show
a change in the IL4R mRNA expression on IL-4 stimulation
(Fig 3A). However, the IL-4R was functional because IL4-stimulated cells showed an increased CD14 antigen expression of 40%, whereas the HLA-DR antigen expression
decreased to 30% after IL-4 stimulation. To exclude the
possibility that the unresponsiveness of the IL-4R for IL-4
was related to the used cell line, monocytes were stimulated
with IL-4. Asdepicted in Fig 3B, no change in IL-4R mRNA
expression was noticed. In three separate experiments,
monocytes showed no significant increase in IL-4R mRNA
expression on stimulation with IL-4 for 6 hours (1.01-fold
2 0.09-fold increase compared with unstimulated monocytes
(mean 2 SEM; n = 3)).
Analysis of IL-4R transcription rates. Run-on assays
were performed to determine whether the effect of PMA and
DBcAMP involves an increase in the transcription rate of
the IL-4R gene. A total of 50 X IO" Mono Mac 6 cells were
stimulated with medium, PMA, and DBcAMP for 5 hours.
Unstimulated Mono Mac 6 cells showed a spontaneous transcription of the IL4R gene, which is in accordance with the
constitutive mRNA expression of the IL4R. In three separate experiments, PMA- and DBcAMP-stimulated cells
showed a 1.33-fold 2 0.13-fold and I .81-fold f 0.32-fold
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DE WIT ET AL
612
k
Q)
a
a
lu
rl
a
Q
T cells Mono Mac 6
Cycles
YC
GAPDH
Fig 6. Detection of ye m R N A using RT-PCR with totalcellular RNA
of human T cells and Mono Mac 6 cells. Specific primer pairs were
used to detect y. m R N A expression (resulting in a 564-nucleotide
fragment) and the GAPDH m R N A expression (resultingin a 429-nucleotide fragment) for equal amplification.
increase in the transcription rate. respectively, compared
withthat for unstimulated cells (mean 2 SEM; n = 3). A
representative experiment is shown in Fig 4. These data
indicate that transcriptional mechanisms are involved in the
upregulation of IL-4R mRNA after stimulation of PKC- and
PKA-dependent pathways.
Analysis of IL-4R InRNA stahilih. Mono Mac 6 cells
were stimulated withPMA or DBcAMP for 6 hours.and
RNA synthesis was blocked by the addition of actinomycinD ( I O yglmL). At different time points, total cellular RNA
was isolated and analyzed for IL-4R mRNA expression. Unstimulated cells showed an IL-4R mRNA half-life of approximately 120 minutes (Fig 5). On activation with PMA, IL-4R
mRNA was not stabilized. However, in DBcAMP-stimulated
Mono Mac 6 cells, IL-4R mRNA was slightly stabilized and
showed a half-life of 1 S0 minutes. The data indicate that the
accumulation of IL-4RmRNA after stimulation of PKAdependent pathways involves stabilization at the posttranscriptional level.
Anal.vsis of the common y c expression. Because the common yc is an essential component of the IL-4R complex in
T cells, we questioned whether the yc is also expressed in
monocytic cells. Total cellular RNA from T cells and Mono
Mac 6 cells was analyzed with RT-PCR. Figure 6 shows a
significant expression of the common yc in T cells, whereas
in Mono Mac 6 cells no transcripts could be detected even
after 40 cycles of amplification. The difference could not be
ascribed to an insufficient amount ofRNA in view of the
results obtained with the GAPDH primers.
Analysis cf IL-4R protein. To determine whether the
changes in IL-4R expression at mRNA levelcorrespond with
changes in 1L-4R protein expression, Mono Mac 6 cells were
stimulated with PMA or DBcAMP for 48 hours, and IL-4R
expression was analyzed using an MoAb. Figure 7 shows
that unstimulated (control) Mono Mac 6 cells express IL-4R
on the cell surface. PMA stimulation results in an increased
number of cells expressing IL-4R (Fig 7A) thatcould be
further augmented by DBcAMP (Fig 7B).
Subsequently, CD23 antigen expression was studied with
the focus on whether the upregulation of IL-4R in response
to PMA stimulation might affect the 1L-4 response. Mono
Mac 6 cells cultured with IL-4 alone showed a 1.90-fold 2
0.08-fold increase in CD23 expression compared with that
of unstimulated cells, whereas a 1.20-fold 2 0.03-fold increase was noticed in response to PMA. However, prestimulation with PMA followed by IL-4 during 24 hours resulted
in a 3.24-fold 5 0.3-fold increase in CD23 expression (mean
2 SEM; n = 3).
DISCUSSION
In the present study, we analyzed the involvement of IL4 and different intracellular signaling pathways in the regulation of IL-4Rs on human monocytic cells. The results show
an upregulation of the IL-4R on activation of the PKC- and
c-AMP-dependent pathways, whereas stimulation with IL-4
or the calcium ionophore A23 I87 did not affect the receptor
expression. These findings suggest that the control of IL-4R
expression on monocytic cells is more restricted than that
on T cells because T cells show an augmented IL4R mRNA
expression on stimulation withPMA,A23187,
anti-CD3,
concanavalin A, and IL-4.17~'"
A remarkable observation was
the fact that PMA did augment the receptor expression,
whereas no effect of IL-4was observed. The unresponsiveness to IL-4 was not caused by a lackof functional
receptors because IL-4 modulated the CD14 and CD23 antigen expression. Arruda and Ho'" recently observed that IL4-exposed monocytes showed a significant redistribution of
PKC activity from cytosol to the nuclear fraction, indicating
that PKC-dependent signaling is triggered on IL-4 stimulation. The apparent discrepancy between the IL-4-mediated
signaling and PMA may be connected to activation of different PKC isoforms on stimulation with one of the activators.
Alternatively, the composition of the IL-4R may be different
between monocytic cells and T and B cells because a strong
increment in IL-4R expression is noticed in both lymphoid
cells in response to IL-4 stimulation.'*.'' Recently performed
studies have shown that binding of IL-4 to the TF-I cell line
can be competed for by 1L-13, suggesting that anIL-4R
complex maybe composed of different subunitszx as has
been shown for IL-3/IL-5 and IL-7 receptor families.'""' In
fact, the IL-4R shares the common yc withthe1L-2Rand
IL-7R."'"2 However, in the Mono Mac 6 cell line, yc transcripts were not detected with RT-PCR, whereas in T cells
a significant expression was noticed. This is in accordance
with findings of Takeshita et al" who could only detect yc
mRNA in lymphoid cells, whereas in the monocytic cell line
THP-I, yc mRNA was absent. The absence of the yc might
affect the IL-4 response in T cells and monocytic cells. This
is supported by findings of Russell et allz who showed tyro-
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613
IL-4R REGULATION IN MONOCYTIC CELLS
Fig 7. Analysis oflL-4R expression onthe call surface of Mono Mac 6 cells stimulated with medium
moll
(control), PMA (50nglmL), and DBcAMP
L) for 48 hours. Cells were labeled with a mouseantihuman IL4R MoAb and analyzed on a FACS.
sine phosphorylation of the insulin receptor substrate-l (IRSl) in response to IL-4 in murine L cells, which constitutively
express the IL-4R and were transfected with the y c . L cells
that were not transfected with the yc did not phosphorylate
IRS-1 in response to IL-4 stimulation, indicating that yc
expression is essential for at least some IL-4-mediated signaling events.
Previously, we showed that monocytes incubated with
PMA for 1 hour showed a reduced binding of I'25-IL-4,34
whereas in the present study, an augmented IL4R expression
was noticed after a longer period of stimulation. The findings
mayreflect different cellular processes in a sequence of
events. The primary response to PKC activation is a downregulation of the IL4R as result of shedding or receptor
internalization, which is followed by a protein-dependent
upregulation of the receptor. Comparable observations have
been reported for the granulocyte-macrophage CSF (GMCSF) receptor with regard to stimulation with GM-CSF and
pMA.35-37
Little has been reported regarding the regulation of IL4R expression on myeloidhonocytic cells. Feldman et aI3*
showed an increment in IL4R expression on the murine
myeloid leukemia M1 cells in response to IL-6 stimulation,
whereas interferon-y did not modulate the receptor expression. In contrast, interferon-y-stimulated murine macrophage-like cells 5774.16 showed an increased IL-4R expression." Studies with additional physiological stimuli will
Controllunlabeled
further reveal the exact regulation of IL-4Rs on monocytic
cells.
A profound effect on IL4R mRNA accumulation was
observed by DBcAMP and PGE2, whichwas caused by
changes at transcriptional and posttranscriptional level in
parallel withan increased IL-4R protein expression. The
CAMP-dependentpathway seems to be of importance for the
signal transduction in monocytic cells, because it affects the
expression of different cytokines and receptors. An increased
expression of tumor necrosis factor receptors has been observed in monocytic cells in response to stimulation with
DBcAMP.4' In addition, CAMP augments the expression of
IL-l and IL-6 at mRNA and protein le~e1,""~'whereas MCSF is downregulated in activated monocytic cells.42The
modulating effects of CAMP on the IL4R expression might
be an important feedback mechanism during an inflammatory response. High concentrations of PGE2can be produced
locally by monocytic cells in response to endotoxin stimulat i ~ and
n ~result
~ in the induction of different cytokines, such
as IL-1, IL-6, and IL-8, by direct and paracrine pathways.
Because of the interactive process between T cells and monocytes, the secreted IL-4 will downregulate the production of
the different ~ytokines."~~
In addition, the controlling effects
of IL-4 on monocytic cells are sustained during the inflammatory response as result ofan upregulation of IL-4R on
activation of CAMP-dependent pathways.
In summary, the data show a PKA- and PKC-dependent
From www.bloodjournal.org by guest on October 21, 2014. For personal use only.
DE WIT ET AL
614
regulation of I L 4 R in monocytic cells, whereas stimulation
of Caz'-dependent pathways and stimulation with
IL-4 had
no effect. I L 4 R regulation in monocytic cells seems to
be
different from the regulatory mechanisms in human T lymphocytes that might relate to the expression of different IL4R components.
ACKNOWLEDGMENT
We thank Dr M.K. Spriggs (Immunex) and Dr S.C. Clark (Genetics Institute) for providing the L-4R and M-CSF cDNA probes. We
are grateful toDr P. Trotta (Schering Plough) for providing IL-4
and to Dr S. Gillis (Immunex) for providing the IL-4R MoAb. The
Mono Mac 6 cell line was kindly provided by Dr H.W.L. ZieglerHeitbrock (Heidelberg, Germany).
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