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Induction of fibroblast proliferation by interleukin-1 derived from human monocytic leukemia cells.

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INDUCTION OF FIBROBLAST PROLIFERATION BY
INTERLEUKIN-1 DERIVED FROM HUMAN
MONOCYTIC LEUKEMIA CELLS
ARNOLD E. POSTLETHWAITE, LAWRENCE B. LACHMAN, and ANDREW H. KANG
Human interleukin-1 (IL-l), free of contaminating lymphokines, was isolated from cultures of purified
monoblasts from a patient with acute monocytic leukemia. Partially purified IL-1 (diafiltration, ultrafiltration, and isoelectric focusing) stimulated proliferation of
subconfluent human fibroblasts in vitro. Further purification of IL-1 by high-resolution gel filtration- and
anion exchangehigh performance liquid chromatography revealed that fibroblast proliferation activity could
not be separated from IL-1 activity (thymocyte proliferation), suggesting that both activities are the properties of a single molecule. Fibroblasts and thymocytes
exhibited a similar sensitivity to the proliferative effects
of IL-1. These findings suggest that macrophages participating in inflammatory reactions in vivo might release
IL-1, which could function to expand fibroblast populations a t sites of inflammatory reactions, by acting as a
fibroblast growth factor.
Interleukin-1 (IL-l), or lymphocyte activating
factor (LAF), is a monokine released in vitro by
cultured monocytes or macrophages. Interleukin-1
acts on a variety of target cells in a genetically
From the Laboratory of Cellular Immunology and the
Medical Service, Veterans Administration Medical Center, the
Departments of Medicine and Biochemistry, University of 'l'ennessee Center for the Health Sciences, Memphis and the Department of
Cell Biology, M. D. Anderson Hospital, Houston, Texas.
Supported in part by NIH grants AM16506 and AM20634
and by the Veterans Administration.
Arnold E. Postlethwaite, MD; Lawrence B. Lachman,
PhD; Andrew H. Kang, MD.
Address reprint requests to A. E. Postlethwaite, MD,
Research Service (151), VA Medical Center, 1030 Jefferson Avenue,
Memphis. TN 38104.
Submitted for publication August 22, 1983; accepted in
revised form March 30, 1984.
Arthritis and Rheumatism, Vol. 27, No. 9 (September 1984)
unrestricted manner. The ability of IL-1 to stimulate
proliferation of mouse thymocytes and synergistically
increase thymocyte response to phytohemagglutinin
or concanavalin A is its most widely recognized property (1,2). Interleukin-1 has recently been demonstrated to affect an ever-increasing number of biologic
systems. Specifically, IL-1 enhances the mitogen response of T cells to lectin and antigen, enhances the
generation of cytotoxic T cells, induces antibody production by B cells, stimulates collagenase and prostaglandin production by rheumatoid synovial cells, and
may affect the systemic response to inflammation by
inducing fever and stimulating the release of acutephase reactants (1,2).
Recent studies have demonstrated that human
peripheral blood mononuclear leukocytes, when stimulated by T cell antigen, mitogen, or allogeneic cells,
produce large (75-60K) and small (16-13K) molecular
weight proteins that stimulate fibroblast proliferation
in vitro (3,4). One of the fibroblast proliferation factors
isolated from supernatants of secondary human allogeneic mixed lymphocyte cultures has been shown to
possess physical and functional properties similar to
human IL-I (4). Partially purified IL-1 induced from
phorbol myristate acetate (PMA)-stimulated murine
P388D1cells has been shown to be capable of stimulating fibroblast proliferation (4).
However, in light of recent data published by
Orosz et a1 ( 9 , caution must be used in interpreting
results observed with PMA-induced IL-1 . To date, no
studies have demonstrated that IL-1 obtained from
cultures of pure populations of cells of the human
monocytic series (devoid of lymphokines) is able to
stimulate fibroblast proliferation. Such a demonstra-
POSTLETHWAITE ET AL
996
tion is crucial before it can be accepted that human IL-1
can trigger fibroblast growth, since it has been previously reported that guinea pig and human T cells, a
human T cell leukemia line (HSB2), and human T cell
lines produce a fibroblast proliferation factor when
stimulated in vitro (6-9). In light of these findings, it is
possible that IL- 1 purified from supernatants obtained
from cultures of lymphocytes and monocytes might be
contaminated with lymphokines, one of which could
stimulate fibroblast proliferation.
The study of human IL-1 has been greatly
facilitated by the recent observation that leukapheresed buffy coat cells from patients with acute monocytic or myelomonocytic leukemia produce large
amounts of IL-1 when cultured in vitro (10). Interleukin-1 from these leukemic cells appears t o be identical
to IL-1 derived from normal human monocytes, and is
not Contaminated by lymphokines (10). In this report,
we have demonstrated that IL-1 purified from supernatants of cultures of cells from a patient with acute
monocytic leukemia is a potent stimulator of fibroblast
proliferation in vitro. This observation suggests that
monocyte-macrophages may have the ability t o stimulate fibroblast growth in vivo by releasing interleukin-1 .
MATERIALS AND METHODS
IL-1 purification. IL- 1 was produced, as previously
described, by stimulating short-term primary cultures of
peripheral blood monoblasts, obtained by leukapheresis of a
patient with acute monocytic leukemia, with lipopolysaccharide 01 11 :B-4 from Eschericlzia coli (Difco, Detroit, MI) (10).
The diagnosis of acute monocytic leukemia was established
by the Hematology Division, Department of Medicine, Duke
University Medical Center, Durham, NC by cytochemical
staining and morphologic appearance of peripheral blood
and bone marrow cells. All of the peripheral blood white
cells isolated from this patient stained positively for nonspecific esterases, and 99% were morphologically monoblasts.
For some studies, IL-1 was produced by stimulating normal
human peripheral blood leukocytcs with lipopolysaccharide
Olll:B-4 (11).
Harvested supernatants containing IL-1 were subjected to diafiltration, ultrafiltration, and isoelectric focusing
(IEF) as previously described (10,12,13). Fractions in the
6.8-7.2 pH range containing IL-1 activity were pooled and
dialyzed at 4°C against large volumes of 0.9% NaCI, and
sterilized by filtration through 0.2-pm Acrodisc filters (Gelman Sciences, Ann Arbor, MI). These purification steps resulted in the separation of lipopolysaccharide from IL-I . IEFpurified IL-I contained background levels of endotoxin (<10
ng/ml) as determined by the Limulits lysate clot assay (14).
High performance liquid chromatography. IL- 1 isolated by IEF from supernatants of cultures of cells from the
patient with monocytic leukemia was found to contain many
contaminating proteins when subjected to sodium dodecyl
sulfate-polyacrylamide gel electrophoresis (14). Therefore,
IEF-purified IL-l was subjected to high performance liquid
chromatography (HPLC) to obtain more highly purified IL-I
and to determine whether its fibroblast and thymocyte
proliferation activities could be separated.
Gel filtration-HPLC was performed on 2 1-125 protein analysis columns (Waters Associates, Milford, MA)
connected in tandem. The columns were equilibrated and
run with 0.0075M glycylglycine/O.14M NaCl (GGBS) at pH
7.2. Analytical anion exchange-HPLC was also performed
on IEF-purified IL-1 from the monocytic leukemia cells. The
anion exchange column (250 x 4.1 mm, SynChropak AX300,
SynChrom, Linden, IN) and IEF-purified IL-1 were equilibrated with starting buffer (0.02M Tris acetate, pH 8).
Samples applied to the column were eluted with a linear salt
gradient established by mixing increasing amounts of limiting buffer (0.02M Tris acetatel0.5M sodium acetate, pH 8)
with starting buffer. The gradient was run so that a 100%
concentration of limiting buffer passed through the column
30 minutes after application of the sample.
Thymocyte proliferation assay. Interleukin- 1 activity
was quantitated by measuring the uptake of tritiated thymidine (3HTdR) by thymocytes from 6-10-week-old SwissWebster mice (Charles River, Boston, MA) (10).
Fibroblast proliferation assay. Primary cultures were
established from explants of human infant foreskins by
standard techniques. Eagle's minimum essential medium
supplemented with nonessential amino acids, ascorbic acid
(50 pg/ml), NaHC03 and HEPES buffer (pH 7.2), penicillin
(100 unitdml), streptomycin (100 pg/ml), amphotericin B (1
pg/ml), and 10% heat-inactivated fetal calf serum (FCS) was
used as maintenance medium.
Fibroblasts in their fourth to twelfth subpassage were
harvested from stock cultures by trypsinization and suspended in maintenance medium at a density of 5 x lo4
cells/ml (15). Aliquots (SO0 pl) were dispensed into wells of
Falcon 3008 multiwell plates (Falcon Plastics, Oxnard, CA)
and cultured for 16 hours at 37°C in a humidified atmosphere
containing 5% COz. Culture medium was then aspirated
from each well and replaced with 500 pI serum-free maintenance medium. Four hours later, medium was removed from
each well and replaced with fresh, serum-free maintenance
medium (300 pl) and aliquots (200 p1) of samples being tested
for proliferation activity. Samples were tested in triplicate.
Prepared plates were routinely incubated for 68 hours at
37°C in a humidified atmosphere containing 5% COz.
Preliminary studies revealed that maximal difference
in 'HTdR uptake between control and IL-I-stimulated fibroblasts could be demonstrated by pulsing cultures at 68 hours
and harvesting 4 hours later. Therefore, 3HTdR (1 .O pCi/2S
pl, specific activity 1.9 Ci/mmole, Amersham, Arlington
Heights, IL) was added to each well at 68 hours. Four hours
later, fibroblast wells were washed 3 times with GGBS, airdried, and coated with clear lacquer. Well bottoms, removed
from the plates by using a punch, were placed in scintillation
vials containing 10 ml Aquasol (New England Nuclear,
Boston, MA). The plastic well bottoms wcre dissolved by
the Aquasol and exerted no detectable quenching effect on
tritium. Incorporated 'HTdR was quantitated in a scintillation counter. Final proliferation activity was expressed,
where indicated, as the mean of the replicates. Standard
error of the mean of triplicate determinations was less than
10%.
997
IL-1 STIMULATION OF FIBROBLASTS
Table 1. Validation of the fibroblast proliferation assay
Quantitation of fibroblast growtht
Fibroblast cultures*
Without FCS
Saline
IL-1 (1:500)
With 5% FCS
Saline
1L-l (1500)
Proliferation assay
(mean cpm ? SE)
*
'
TCA-precipitated
(mean cpm 2 SE)
127
7
700 + 60
51
564
2.018 ? 158
4,895 t 208
565
1,527
2
2
13
20
2 84
2 4I
Fibroblast numbers
(mean cells/5 LPF)
270
703
*
?
18
42
532 ? 36
1,160 2 112
*FCS = fetal calf serum; IL-1 = interleukin-I. See text for culture details.
tTCA = trichloroacetic acid; LPF = low-power field. See text for assay details.
RESULTS
Validation of fibroblast proliferation assay. It
was essential to establish that the fibroblast proliferation assay measured actual fibroblast growth in response to IL-1. To accomplish this, fibroblasts in
maintenance medium containing 10% FCS were added
at a density of 2.5 x lo4 cells/well to 3 multiwell plates.
Medium was changed to serum-free maintenance medium. Four hours later, it was replaced with serumfree maintenance medium or maintenance medium
containing 5% FCS. Saline or IEF-purified IL-1 from
normal human monocytes was added to triplicate wells
of each plate (1500 final dilution). After 68 hours
incubation, plates 1 and 2 were pulsed with 3HTdR,
and 4 hours later they were washed 3 times with
GGBS. Plate 1 was air-dried and well bottoms were
removed and placed in vials containing Aquasol (10
ml) for scintillation counting. The cell layers on plate 2
were solubilized by 0.1% sodium dodecyl sulfate (50
pl/well). Carrier protein (1:20 diluted guinea pig serum, 50 pl/well) was added to each well. Cellular
protein, DNA, and carrier protein were then precipitated with cold 10% trichloroacetic acid (TCA). TCA
precipitates were collected on glass paper filters
placed in a vacuum device. Filters were air-dried
overnight and placed in vials containing Aquasol (10
ml) for scintillation counting. In plate 3 , fibroblast
numbers were determined after culturing for 5 days.
Fibroblast nuclei were counted in 5 random fields in
each well at 2 0 0 ~magnification.
As illustrated in Table 1, IL-1 stimulated both
the incorporation of 'HTdR into fibroblast DNA and
an actual increase in the number of fibroblasts in
serum-free and serum-containing cultures. As expected, serum-containing cultures to which saline or IL-I
was added had higher counts. By all 3 methods of
quantitation, serum-free cultures exhibited a greater
percentage increase in stimulation in response to IL-1
than cultures containing 5% FCS (Table 1).
It was also of interest to determine whether the
fibroblast proliferation assay was valid for confluent
fibroblast cultures. Fibroblasts in maintenance medium containing 10% FCS were added at a density of 5 x
lo4 cells/well to a Falcon 3008 multiwell plate. The
plate was incubated for 3 days, at which time fibroblasts reached confluency. Medium was removed from
each well and replaced with serum-free maintenance
medium. Four hours later, medium was again removed
from each well. To wells in one-half of the plate,
serum-free maintenance medium (400 ml) was added,
and to wells in the other half of the plate, maintenance
medium (400 ml) containing 6.25% FCS was added. To
6 wells in each half of the plate, saline (100 pl) was
added, and to 6 wells in the other half of the plate, IL-1
(1:lOO dilution of the same preparation used in the
experiment described above) was added. After 68
hours additional incubation, each well was pulsed with
1.0 pCi/25 pl of 'HTdR. Four hours after the 3HTdR
pulse, half of the plate wells were harvested by the
protocol used in the fibroblast proliferation assay
described in Materials and Methods, and the other half
of the plate was harvested by TCA precipitation as in
the experiment described above.
As shown in Table 2, IL-1 caused a twofold
increase in 3HTdR uptake by confluent fibroblasts in
the presence of 5% FCS. Parallel cultures harvested
by TCA precipitation revealed no increase in 3HTdR
incorporation into nuclear DNA in IL-l-containing
cultures supplemented with 5% FCS. However, serum-free confluent fibroblast cultures harvested by
both techniques did not show increased 3HTdR uptake
or incorporation into DNA.
These data suggest that the fibroblast proliferation assay, which measures both 3HTdR uptake by the
cell and incorporation into DNA, is a valid measure of
POSTLETHWAITE ET AL
998
Table 2. Effect of IL-I on growth of confluent fibroblast cultures
Quantitation of fibroblast growtht
Fibroblast cultures*
Without FCS
Saline
1L-1 (1:SOO)
With 5% FCS
Saline
IL-1 f1:SOO)
Proliferation assay
(mean cpm SE)
*
TCA-precipitated
(mean cpm SE)
*
23
10
952 6
1,328 ? 71
2.644 ? 132
842 2 67
731 ? 16
250
260
2
?
89?
7
* FCS = fetal calf serum; IL-I = interleukin-I. See text for culture
details.
t TCA = trichloroacetic acid. See text for assay details.
fibroblast growth for subconfluent fibroblast cultures
with or without FCS supplementation, but it is a valid
measure of fibroblast growth in confluent fibroblast
cultures only if they contain no serum. These data and
data we have previously published suggest that IL-1
can stimulate replication of subconfluent fibroblasts
but does not stimulate replication of confluent fibroblasts (16).
Relative sensitivity of fibroblasts and thymocytes
to the proliferative effect of IL-1. IL-I isolated by
diafiltration, ultrafiltration, and isoelectric focusing of
supernatant from cultures of lipopolysaccharide-stimulated normal human peripheral blood mononulcear
leukocytes was also found to contain a contaminating
lymphokine, lymphocyte-derived chemotactic factor
for monocytes (chemotactic activity of the IL-1 preparation was 54 6 monocytes per oil immersion field
[OIF], compared with a buffer control of 5 ? 1
monocytes per OIF).
Since a human T cell-derived factor has also
been found to be capable of stimulating fibroblast
proliferation, we isolated IL-1 from cultures of monoblasts (99% pure) of a patient with acute monocytic
leukemia. All of this patient's white cells stained
positively for nonspecific esterases (17). We found
that IEF-purified IL- 1 isolated from supernatants of
cultures of monoblasts from this patient did not contain this lymphokine activity (chemotactic activity of
this IL-1 preparation was 6
1 monocytes per OIF
compared with a buffer control of 7 ? 1 monocytes per
OIF). Since it was desirable to obtain IL-1 free of
contaminating lymphokines, we used supernatants
from cultures of monoblasts and monocytes from this
patient as our primary source of IL-1. IL-I from this
patient was used in all subsequent experiments.
It was of interest to assess the relative sensitivity of fibroblasts and thymocytes to the proliferative
effects of IL-1; therefore, different doses of IEF-
*
*
purified IL-1 were tested for their ability to stimulate
thymocyte and fibroblast proliferation. We performed
such dose-response experiments on 3 occasions. As
illustrated by a representative experiment shown in
Figure 1, the proliferative response of fibroblasts paralleled that of thymocytes and was linear with the
doses of IL-1 tested. This suggests that fibroblasts and
thymocytes have a similar sensitivity to the proliferative effect of IL-1.
Coelution of IL-1 and fibroblast proliferation
activities from HPLC columns. Since IEF-purified IL- 1
from monocytic leukemia cells was not homogenous,
it was possible that it contained a different monokine
responsible for the stimulation of fibroblast proliferation. In an effort to answer this question, we subjected
IL-I to further analysis by high-resolution gel filtration- and anion exchange-HPLC on 3 separate occasions. Thymocyte and fibroblast proliferation assays
were performed on the HPLC column fractions. Fibroblast proliferation and IL- I activities consistently coeluted from 1-125 gel filtration-HPLC columns in a
single major peak, as illustrated by the experiment in
Figure 2. When IEF-purified IL-I was applied to the
1000~
I\
-4000
900-
2
3
-3500 X
aoo-
-3000
:
.5-
U
C
2
0
T
z
-2500
700-2000
E
--
%
3
.-c0
F
X
y
Y
0
6001500
n
5
.
D
m
n
g 500-
1000
0
m"
LL
t
4 o o0j
500
I L 1 Dilution
Figure 1. Fibroblast and thymocyte proliferation assays. Human
interleukin-I (IL 1) was partially purified (diafiltration, ultrafiltration, and isoelectric focusing) from supernatants of cultures of
monoblast-monocytes from a patient with acute monocytic leukemia
and tested at the dilutions indicated.
999
IL-1 STIMULATION O F FIBROBLASTS
mLL
04
0
20
30
40
50
I0
60
I
u
c
FRACTION NUMBER
Figure 2. Fibroblast and thymocyte proliferation assays. A sample
(0.3 ml) of isoelectric focusing-purified interleukin- 1 was applied to
2 1-125 protein analysis columns connected in tandem. Columns
were run at a Row rate of I ml/minute. Fractions (0.4 ml) were
collected and tested at a 1:6 dilution.
mechanisms by which IL-1 exerts mitogenic effects on
these target cells are apparently different. Studies by
Smith et al(l8) suggest that IL-I stimulates a subpopulation of thymocytes to release IL-2, which in turn
binds to a receptor on a larger population of thymocytes, causing them to undergo mitogenesis. IL-2 is
not involved in IL-1 triggering of fibroblast mitogenesis. Schmidt et a1 (4) have clearly shown that IL-2 is
not mitogenic for fibroblasts. In addition, the monocytic leukemia IL-1 used in this study contained no
IL-2 activity (Lachman L: personal observation). The
mechanisms involved in IL- 1 stimulation of fibroblast
growth remain to be defined.
Recent reports have shown that in addition to
IL-1, there is a lymphocyte-derived factor capable of
stimulating fibroblast proliferation (7-9). Such a factor
may be present in supernatants obtained from secondary human allogeneic mixed lymphocyte reactions
which were found, by Schmidt et al (4), to contain a
fibroblast growth factor. It is possible that IL-1 was
responsible, in part, for the fibroblast proliferation
activity characterized in that rcport. Murine IL- 1 was
shown in that same study to stimulate fibroblast prolif-
anion exchange-HPLC column, fibroblast proliferation and IL-1 activities were always confined to a
single peak that eluted from the column between 21
and 25% of the salt gradient, as illustrated in Figure 3.
These data obtained by the high resolution afforded by
gel filtration- and anion exchange-HPLC strongly
suggest that IL-1 is capable of acting as a mitogenic
stimulus for both fibroblasts and thymocytes.
DISCUSSION
Human IL-1 produced by monocytic leukemia
cells and purified by diafiltration, ultrafiltration, and
isoelectric focusing stimulated subconfluent dermal
fibroblasts to proliferate in a dose-dependent manner.
Fibroblasts and thymocytes exhibited a similar sensitivity to IL-1. Further analysis of IEF-purified IL-1 by
high-resolution gel filtration- and anion exchangeHPLC revealed that thymocyte and fibroblast proliferation activities could not be separated from each
other. These data strongly suggest that IL-1 can trigger
not only the proliferation of thymocytes, but the
growth of fibroblasts as well.
Although the same doses of IL-1 that stimulate
thymocyte proliferation also trigger fibroblast growth,
100
0
0
0.041
10
20
30
40
50
60
-rlW
30
40
50
60
.
0
10
20
FRACTION NUMBER
Figure 3. Fibroblast and thymocyte proliferation assays. A sample
(0.3 ml) of isoelectric focusing-purified interleukin-I was applied to
a SynChropak AX300 anion exchange column and eluted with a
linear salt gradient at a flow rate of 1 m h i n u t e . Fractions (0.5 ml)
were collected and tested at a 1:6 dilution.
POSTLETHWAITE ET AL
eration (4). However, the results are less clear, since
PMA was used to induce IL-1 production.
It has recently been shown that PMA can form
complexes with elements in tissue culture medium and
interfere with a variety of 1L-1 bioassays by mimicking
various IL-1 effects (5). Our present study has circumvented the problem of lymphokine cantamination of
IL-1 preparations by using as a source of IL-1 a
population of monoblasts and monocytes containing
no lymphocytes, obtained from a patient with acute
monocytic leukemia. In addition, the problem of PMA
contamination was avoided since this patient’s monoblasts produced 1L-1 in response to endotoxin.
Several different reports have shown that cells
of the mononuclear phagacyte series elaborate a factor(s) that stimulates growth of fibroblasts (19-24).
However, similarity of these factors to IL-1 has not
been demonstrated previously. Only one report, by
Bitterman et a1 (24), has differentiated these factors
from IL-1. They found that normal human alveqlar
macrophages, when stimulated in suspension culture
by opsonized zymosan, opsonized Sepharose, heatkilled Strepromyces albus, IgG-immune complexes,
IgM-complement-immune complexes, or by attachment to plastic, elaborated a factor with a molecular
weight of about 18,000, that stimulated growth of
“noncycling” human lung fibroblasts (24). This factor,
when partially purified, did not stimulate thymocyte
mitogenesis. Interestingly, in their assay, which used
established embryonic lung fibroblasts or adult lung
fibroblasts as target cells, partially purified human IL1 did not trigger fibroblast replication. In performing
their assay, they grew fibroblast targets in medium
containing 0.4% FCS for 2 days and added plateletderived growth factor for 3 additional days before
adding IL-1. Incorporation of ’HTdR was measured
only during the 24 hours after addition of IL-1. The
target cells they used (lung fibroblasts, compared with
dermal fibroblasts in this study) and many other conditions under which they performed the fibroblast proliferation assay (24) differed markedly from those used in
the present study.
Numerous substances in addition to endotoxin
have been reported to stimulate IL-1 release from
macrophages in vitro. Specifically, PMA, muramyl
dipeptide, lymphokines, phagocytosed bacteria, and
particulates have been shown to lead to IL-1 release
by macrophages, suggesting that IL-1 might be released by macrophages participating in inflammatory
reactions of diverse etiologies (1). These studies must
be interpreted with caution because they were not
controlled for endotoxin contamination. Endotoxin is
an extremely potent stimulator of IL-1 production by
macrophages (13).
Regulation of IL-1 release in vivo has not been
studied in depth. However, several studies have documented the presence of IL-1 in inflammatory reactions
associated with accelerated fibroblast accumulation.
IL-1 and additional fibroblast growth factors have
been isolated from murine schistosomal granulomas
(25). An IL-1-like factor has recently been reported to
be present in synovial fluid of patients with different
types of arthritis (26).
It is becoming increasingly apparent that the
fibroblast is an important target cell for IL- 1. Human
IL-1 not only stimulates growth of subconfluent fibroblasts in vitro, but we have recently reported that it
also stimulates the production of interstitial collagenase by cultures of confluent dermal fibroblasts (16).
This stimulation of collagenase production by cultures
of confluent fibroblasts is not associated with increased DNA synthesis (25). Production of collagenase by rheumatoid synovial cells, which show some
similarities to fibroblasts, is also stimulated by IL-1 or
a closely-related monokine (27).
The ability of 1L-1 and other monokines to
serve as fibroblast growth factors may have importance in vivo. Fibroblasts characteristically appear at
sites of inflammatory reactions after an influx of
macrophages and repair tissue damage by synthesizing
new matrix components. We have previously suggested that the expansion of fibroblast populations at sites
of inflammatory reactions may result in part from
migration of these cells in response to specific chemoattractants (15,28-31). This study suggests that
macrophages may also be capable of expanding fibroblast populations at sites of inflammatory reactions by
releasing IL-1, which in turn acts as a mitogen for
fibroblasts.
ACKNOWLEDGMENTS
The authors acknowledge the excellent technical
assistance of Susan Turner and Patricia Dean and the
secretarial assistance of Dorothy K. Davis. The technical
suggestions by Dr. Jerome M. Seyer were most useful in
carrying out HPLC analyses.
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induction, leukemia, monocytic, proliferation, interleukin, derived, human, cells, fibroblasts
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