| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
|
|
|
|||||||
|
|||||||||
*
Department of Biochemical Pharmacology, The William Harvey Research Institute, London, United Kingdom; and
Division of Critical Care Medicine, Children’s Hospital Medical Center, Cincinnati, OH 45229
 |
Abstract |
|---|
 Top Abstract IntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
) and mast cells
(MCs) in polymorphonuclear leukocyte (PMN) accumulation and chemokine
production within the mouse peritoneal cavity in response to
administration of zymosan (0.2 and 1 mg), LPS (1 mg/kg), and
thioglycolate (0.5 ml of a 3% suspension) were investigated. A marked
reduction (>95%) in intact MC numbers was obtained by pretreatment
with the MC activator compound 48/80, whereas resident M were
greatly diminished (>85%) by a 3-day treatment with liposomes
encapsulating the cytotoxic drug dichloromethylene-bisphosphonate. No
modulation of thioglycolate-induced inflammation was seen with either
pretreatment. Removal of either MCs or M attenuated LPS-induced PMN
extravasation without affecting the levels of the chemokines murine
monocyte chemoattractant protein-1 and KC measured in the lavage
fluids. In contrast, MC depletion inhibited PMN accumulation and murine
monocyte chemoattractant protein-1 and KC production in the zymosan
peritonitis model. Removal of M augmented the accumulation of PMN
elicited by the latter stimulus. This was due to an inhibitory action
of M-derived IL-10 because there was 1) a time-dependent release of
IL-10 in the zymosan exudates; 2) a reduction in IL-10 levels following
M, but not MC, depletion; and 3) an increased PMN influx and
chemokine production in IL-10 knockout mice. In conclusion, we propose
a stimulus-dependent role of resident MCs in chemokine production and
the existence of a regulatory loop between endogenous IL-10 and the
chemokine-mediated cellular component of acute
inflammation. |
Introduction |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
promotes PMN and mononuclear cell
recruitment (for a recent review, see 2 .
Many studies have focused on the role played by chemokines in
attracting different subsets of leukocytes in human inflammatory
pathologies and in their experimental counterparts. Experimentally,
several observations have highlighted the strict relationship between
chemokine expression and the intensity of the host inflammatory
response as well as their direct link with the appearance of
blood-derived leukocytes at the site of inflammation. For example, IL-8
is generated in a rabbit model of joint articular inflammation and
promotes PMN extravasation (3). In the mouse, chemokines such as
MCP-1, MIP-2, and MIP-1 contribute to the initiation of articular
arthritis and recruit monocytes and lymphocytes (4). A similar link
between chemokines and leukocyte recruitment also exists in acute
models of inflammation. We have recently shown that MCP-1, MIP-1,
and KC are expressed at early time points (<4 h) in a murine
model of acute peritonitis (5). This is not totally surprising, as some
of the genes that produce chemokines were initially identified as
early/intermediate response genes (e.g., JE in the mouse, which
produces murine (m) MCP-1) (6).
In the present study we use several distinct models of acute
inflammation to address the question of which cell type(s) produces
these chemokines. Because of the early expression of these mediators,
we addressed our attention to M and MCs, since both are resident
cells in close contact with postcapillary venules (7). As demonstrated
by selective depletion experiments, M and MCs produce the initial
signals responsible for the accumulation of PMNs, eosinophils, and
mononuclear cells in experimental models of inflammation (8, 9, 10).
Here, we have 1) monitored the effect of selective removal of
either resident cell type on KC (a murine CXC chemokine) and mMCP-1 (CC
chemokine) gene and protein expression in the mouse and 2) assessed how
their expression is functionally linked to leukocyte migration.
Finally, we describe a novel chemokine/cytokine inhibitory feedback
loop based upon IL-10 released from the M.
|
Materials and Methods |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
Male Swiss Albino mice were purchased from Banton and Kingsman (Hull, U.K.). Male C57BL/6 IL-10 wt and C57BL/6 IL-10 KO were obtained from The Jackson Laboratory (Bay Harbor, ME). The animals were fed a standard chow pellet diet and had free access to water. All animals were maintained on a 12-h light, 12-h dark cycle and housed for 1 wk before experimentation. Mice weighed between 26–30 g on the day of the experiment.
Models of inflammation
Zymosan peritonitis was induced as previously reported (5). Briefly, mice were injected i.p. with zymosan A (0.2 or 1 mg in 0.5 ml of saline). At selected time points, animals were euthanized by carbon dioxide exposure, and peritoneal cavities were lavaged with 3 ml of PBS containing 3 mM EDTA. Aliquots of the lavage fluid were then stained with Turk’s solution (0.01% crystal violet in 3% acetic acid), and differential cell counts were performed using a Neubauer hemocytometer and a light microscope (B061, Olympus, Melville, NY). The lavage fluids were centrifuged at 400 x g for 10 min, and cell-free supernatants were stored at -20°C before chemokine and cytokine evaluation by ELISA (see below). Cell pellets were then prepared for RNA extraction and RT-PCR analysis as described below.
LPS peritonitis was induced as previously reported (5). Mice received i.p. injections of LPS (1 mg/kg/10 ml of sterile saline). At selected time points, peritoneal cavities were washed, and the lavage fluid was handled as described above.
Thioglycolate (TG) peritonitis was induced as previously reported (11). Mice received i.p. injections of TG (3% (w/v) in 0.5 ml of saline). At designated time points, peritoneal cavities were washed, and lavage fluid was handled as described above.
Chemokines and cytokine ELISA
Immunoreactive mMCP-1, IL-10, TNF-, and KC were quantified
using a commercially available ELISA according to the manufacturer’s
protocol. In brief, lavage fluids (100 µl) were assayed for mMCP-1
and compared with a standard curve constructed with 0–2.5 ng/ml murine
mMCP-1. Similarly, lavage fluids were tested for the murine chemokines
KC (standard curve ranging from 0–1 ng/ml) and the cytokines TNF-
(standard curve ranging from 0–1.5 ng/ml) and IL-10 (standard curve
ranging from 0–1 ng/ml). The ELISA method consistently detected KC,
TNF-, and murine IL-10 at >1.5 pg/ml and mMCP-1 at >9 pg/ml. The
ELISAs showed negligible (<1%) cross-reactivity with several murine
cytokines and chemokines (data furnished by the manufacturer).
Detection of chemokine mRNA by RT-PCR analysis
Peritoneal cell pellets (5 x 106) were lysed in 1 ml of Trizol reagent, and RNA was isolated according to the manufacturer’s protocol. Briefly, RNA was extracted with chloroform and isopropanol. The RNA was precipitated with ethanol, and the pellet was resuspended in diethylpyrocarbonate-treated water. The yield and the purity of the RNA were then estimated spectrophotometrically at 260 and 280 nm. Total RNA (3 µg) was used for the generation of cDNA. PCR amplification reactions were performed on aliquots of the cDNA. For the mMCP-1 target, the primers were 5'-ACT-GAA-GCC-AGC-TCT-CTC-TTC-CTC-3' and 5'-TTC-CTT-CTT-GGG-GTC-AGC-ACA-GAC-3' (forward and reverse), which amplified a fragment 274 bp in length. For the KC target, primers were 5'-GGA-TTC-ACC-TCA-AGA-ACA-TCC-AGA-G-3' and 5'-CAC-CCT-TCT-ACT-AGC-ACA-GTG-GTT-G-3' (forward and reverse), which amplified a fragment 454 bp in length.
All PCR reactions were performed in a final volume of 25 µl using a Hybaid OmniGene thermal cyler (Middlesex, U.K.). For mMCP-1, the PCR profile consisted of one cycle of denaturation at 94°C for 2 min, followed by 35 cycles of denaturation at 94°C (45 s), annealing at 57°C (45 s), and extension at 72°C (30 s). For KC, an initial denaturation was performed at 94°C for 2 min, followed by 35 cycles of denaturation at 94°C (45 s), annealing at 54.7°C (45 s), and extension at 72°C (30 s). Amplification products were visualized by ethidium bromide fluorescence in agarose gels. Bands of the expected sizes were obtained. Images were inverted using the Graphic Converter software (version 2.1; Lemke Software, Peine, Germany) running on a Macintosh Performa 6200 (Apple Computer, Cupertino, CA).
Drug treatments
Liposomes.
Selective depletion of resident peritoneal M was achieved by
pretreatment of mice with multilamellar liposomes containing
dichloromethylene-bisphosphonate (Cl2MDP or clodronate) and
was performed according to a published procedure (12). Briefly, 86 mg
of phosphatidylcholine and 8 mg of cholesterol were dissolved in 10 ml
of chloroform in a sterile round-bottom flask by vacuum rotary
evaporation at 37°C. Cl2MDP (1.89 g dissolved in 10 ml of
sterile PBS) was encapsulated into the preparation of
phosphatidylcholine and cholesterol by gently shaking for 10 min. The
solution was kept for 2 h at room temperature, then sonicated for
3 min at 20°C and kept for an additional 2 h at room
temperature. Free Cl2MDP was removed by three
centrifugation steps (100,000 x g for 30 min at 4°C)
in a Beckman Ultracentrifuge L-80 Optima (Beckman, Palo Alto,
CA). Liposomes, layered on top of the supernatants, were
collected and washed twice by centrifugation. The liposome pellets were
resuspended in 4 ml of sterile PBS. Mice were treated i.p. with 100
µl of liposome preparation for 3 consecutive days; 24 h after
the last injection peritoneal cavities were lavaged, and the number of
macrophages was determined following staining in Turk’s solution. In
another set of experiments, zymosan (0.2 or 1 mg), TG (0.5 ml of 3%
suspension), or LPS (1 mg/kg) was administered i.p. 24 h after the
last liposome administration. Peritoneal cavities were lavaged 4 h
later (for zymosan and TG) or 3 and 24 h later (for LPS) for
chemokine and PMN determinations as described above.
M depletion was also confirmed by FACS analysis. In brief,
peritoneal cells (1 x 106) were stained for 60 min on
ice with 20 µg/ml of rat anti-mouse F4/80 mAb or rat IgG2b
isotype control. After extensive washing, cells were stained with
FITC-conjugated anti-rat IgG for 45 min on ice, extensively washed,
and analyzed by flow cytometry using a FACScan analyzer (Becton
Dickinson, Cowley, U.K.). Distinct populations were based on the
forward and side scatter characteristics, and fluorescence
associated with them was measured in the on FL1 channel.
Compound 48/80. Resident peritoneal MCs were depleted using a well-characterized protocol (8). Mice received a single dose of compound 48/80 (1.2 mg/kg, i.p.) or sterile saline (100 µl i.p.) 72 h before lavaging the peritoneal cavities: the number of intact MCs in the lavage fluids was determined following staining in 200 µl of toluidine dye. In another set of experiments, 72 h post-treatment, mice were injected i.p. with zymosan (1 mg), LPS (1 mg/kg), or TG (0.5 ml of a 3% suspension). Peritoneal cavities were lavaged 4 h later (for zymosan and TG) or 3 and 24 h later (for LPS) for chemokine and PMN determinations as described above.
Measurement of nitrite/nitrate concentration in the lavage fluids
Nitrite/nitrate production, an indicator of nitric oxide synthesis, was measured in lavage fluids as previously described (13). First, nitrate in the peritoneal lavage fluids was reduced to nitrite by incubation with nitrate reductase (670 mU/ml) and NADPH (160 mM) at 37°C for 3 h. The nitrite concentration in the samples was then measured by the Griess reaction, by adding 100 µl of Griess reagent (0.1% naphthalethylenediamine dihydrochloride in H2O and 1% sulfanilamide in 5% concentrated H3PO4 (1:1, v/v)) to 100-µl samples. The OD at 550 nm (OD550) was measured using a Spectramax 250 microplate reader (Molecular Devices, Sunnyvale, CA). Nitrate concentrations were calculated by comparison with OD550 of standard solutions of sodium nitrite prepared in saline solution.
Reagents
Quantikine ELISA kits for murine IL-10, TNF-, and KC were
purchased from R&D Systems (Abingdon, U.K.), whereas the specific
murine mMCP-1 ELISA Cytoscreen was obtained from BioSource
International (Camarillo, CA). F4/80 mAb (clone CI:A3–1) and
FITG-conjugated anti-rat IgG were purchased from Serotec (Oxford,
U.K.). LPS (Escherichia coli serotype 0111:B4),
zymosan A, TG, and all other chemicals were purchased from Sigma
(Poole, U.K.). Clodronate was a generous gift from Boehringer Mannheim
(East Sussex, U.K.).
Reagents for the PCR reaction were purchased from the following companies: mMCP-1 and KC primers from OligoExpress (Middlesex, U.K.), Trizol reagent (lysis buffer for RNA preparation) from Life Technologies (Paisley, U.K.), and Ready-to-Go T-Primed First-Strand Kit and PCR Beads from Pharmacia Biosystem Europe (St. Albans, U.K.).
Statistical analysis
Data are reported as the mean ± SEM of n mice per group, and statistical differences were evaluated by one-way analysis of variance once Bartlett’s test confirmed the homogeneity of the variances. Post-hoc comparisons were made with Bonferroni’s test using Instat software (version 2.04) running on a Macintosh Performa 6200. A threshold value of p < 0.05 was taken as significant.
|
Results |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
In initial experiments the different inflammatory responses to the
three stimuli were characterized. Zymosan injection induced a
time-dependent accumulation of PMN into the cavity, with a maximal cell
influx at the 4 h point (Fig. 1
a). PMN accumulation was
almost resolved by 48 h postzymosan administration. Local
injection of TG caused a significant extravasation of PMN into the
cavity, peaking at 4 h and still elevated above basal levels at
48 h postinjection. In contrast, i.p. injection of LPS induced a
delayed influx of PMN, with a high rate of influx between 6–16 h
(
0.9 x 106 PMN/h). Significant numbers of PMN were
still present in the cavity at 24 h, whereas a sharp decline to
near basal levels was detected by 48 h (Fig. 1a). In
all cases, the large predominance of neutrophils (99%) in the PMN
population was confirmed in cytospin preparations stained with
May-Grunwald and Giesma (data not shown).
|
The level of mMCP-1 in the peritoneal lavage fluid was determined
by a specific ELISA. No mMCP-1 protein was found in basal conditions
(Fig. 1
b). As expected (5), i.p. administration of zymosan
resulted in a rapid release of mMCP-1 into the lavage fluids, with
significant amounts at 2 and 4 h and a return to basal levels by
24 h. A time-dependent release of mMCP-1 was also observed
following i.p. injection of LPS: endogenous mMCP-1 could be detected as
early as 1 h postinjection, peaked at 3 h, and then declined
to near basal levels by 24 h (Fig. 1b). Administration
of TG also induced the release of mMCP-1 protein in the lavage fluids,
with peak production measured at 4 h.
Effect of depletion of resident peritoneal MCs or M
Table I shows that pretreatment of
mice with compound 48/80 reduced the number of intact MCs by 95%.
Similarly, a marked depletion (86% reduction) of resident peritoneal
M was obtained by a 3-day treatment of mice with liposomes. M
depletion was confirmed by the disappearance of F4/80 staining as
assessed by FACS analysis (from 463 ± 36 down to 22 ± 4
mean fluorescence intensity units; n = 6;
p < 0.05).
|
a). However, significant
reductions in the number of resident M produced an unexpected (and
significant) increase in cell extravasation. This was investigated
further (see below). The LPS response was also attenuated in
MC-depleted mice compared with that in control animals. A significant
reduction (80%) in LPS-induced PMN influx at 24 h was obtained
following M depletion (Fig. 2b). PMN accumulation into
the peritoneal cavities measured following local injection of TG was
not modulated by either MC or M depletion (Fig. 2c). For
this reason, the TG-induced inflammatory response was not used any
further.
|
)
and KC (5) induced by zymosan or LPS was then assessed. Analysis of
cell extracts by RT-PCR showed the presence of mMCP-1 and KC mRNAs only
in peritoneal cells from zymosan-treated mice and not in control
PBS-injected animals (Fig. 3,
b and e). A reduction in intact MC numbers
reduced the release of both mMCP-1 and KC by >60% as measured in
zymosan lavage fluids (Fig. 3, a and d). An
apparent reduction in KC and mMCP-1 mRNA was seen in MC-depleted mice.
In contrast, no significant modulation was observed of KC and mMCP-1
protein levels in the lavage fluids (Fig. 3, a and
d) or of mRNA in the cell pellets (data not shown) of
M-depleted animals. Depletion of either resident peritoneal MCs or
M did not alter the amounts of KC and mMCP-1 released by LPS (Fig. 3, c and f). In this set of experiments TNF-
levels were also measured, and again no modulation by M depletion
was found (420 ± 15 and 430 ± 10 pg/cavity in intact and
liposome-pretreated mice, respectively; n = 8; not
significant).
|
Based on the results shown above, we decided to characterize
further the mechanism(s) underlying the unexpected increased influx of
PMN after zymosan injection into M-depleted mice. Fig. 4 compares the PMN accumulation measured
after the injection of 0.2 or 1 mg of zymosan; a twofold increase was
seen in the M-depleted animals treated with the lower dose of
zymosan. Lower concentrations of chemokines were released with 0.2 mg
of zymosan (compare Table II with Fig. 3). No significant change in mMCP-1, but a pronounced reduction in KC
concentrations, between control and M-depleted mice was detected
(Table II).
|
|
a). A marked reduction in
resident M, but not MC, numbers significantly affected the amounts
of IL-10 recovered in the lavage fluids at the 4 h point (Fig. 5b).
|
). Similar increases in PMN
accumulation were induced by 0.2 mg of zymosan in IL-10 KO as well as
IL-10 wt mice depleted of peritoneal M. Depletion of M in IL-10
KO mice resulted in a further increase in PMN influx (Fig. 6a). Zymosan injection released comparable amounts of
chemokines in C57BL/6 wt and Swiss Albino mice. Higher mMCP-1 and KC
levels were measured in the lavage fluids of IL-10 KO mice than in
IL-10 wt mice. Further, the release of mMCP-1 lasted longer in IL-10 KO
animals, with values of 2.0 ± 0.1 and 4.4 ± 0.4 ng/cavity
measured in the 8 h lavage fluids in wt and KO mice, respectively
(n = 7; p < 0.05). No modulation by
peritoneal M of mMCP-1 release in either wt or KO animals was seen
(Fig. 6, b and c).
|
depletion protocol in IL-10 KO mice, and since
recent evidence shows that nitric oxide is released in zymosan
peritonitis (13), we monitored nitrate/nitrite levels as a positive
control. A significant reduction in the concentration of
nitrate/nitrite was measured after M removal, from 10.0 ± 1.8
in intact mice to 6.4 ± 0.7 µM in M-depleted mice
(n = 5; p < 0.05). |
Discussion |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
on
the influx of PMN that is effected via the release of endogenous IL-10.
One of the aims of this study was to identify the potential
contribution by resident MCs and M of the production of selected CXC
and CC chemokines during experimental inflammation. We chose to use
three inflammogens: 1) particles of zymosan, a phagocytic stimulus for
the M and the MC, which also causes complement activation and
produces an intense, but short-lasting, influx of PMN (14); 2) TG
broth, which causes a milder and longer lasting accumulation of PMN
(11); and 3) LPS, a soluble M activator that releases endogenous
cytokines to produce PMN influx (15, 16).
The i.p. injection of TG and zymosan induced a rapid and time-dependent accumulation of PMNs in the peritoneal cavity. PMN influx in response to LPS was delayed but was also time dependent. Injection of these inflammatory agents also led to the coordinated, rapid, and time-dependent production of mMCP-1 in the peritoneal lavage fluid as assessed by ELISA. Secretion of mMCP-1 by these inflammatory stimuli reached a peak 4 h post-zymosan and TG administration and 3 h post-LPS injection; in all cases mMCP-1 decreased rapidly thereafter, indicating a fast clearance of the chemokine from the peritoneal cavity. The fact that mMCP-1 induction by these inflammatory stimuli was rapid and transient is not unusual in in vivo experimental systems (5, 17). The rapidity of mMCP-1 induction can be attributed to the fact that this chemokine is the product of an early activation gene (6), whereas the transient nature of its appearance can be attributed 1) to the ability of mMCP-1 to bind to duffy Ag receptor for chemokines, which has been proposed to function as a sink for chemokines (2); or 2) to the release of endogenous inhibitors that switch off the synthesis of the chemokine. We have previously reported an apparent discrepancy between mMCP-1 protein release in the exudates and gene expression in the peritoneal cell pellet, which suggests the existence of such a control mechanism at the level of mMCP-1 mRNA translation (5).
A marked reduction in resident MCs, achieved using a protocol
well validated in our laboratory (8, 10), attenuated the cellular
response produced by both zymosan and LPS. Resident M numbers were
reduced according to a validated procedure that implies the selective
delivery of a cytotoxic drug by neutral liposomes (12). Liposomes have
been reported to produce an anti-inflammatory action per se (18);
however, this is unlikely to be the case in our experiments because 1)
a general anti-inflammatory effect was not seen (compare the
diverse effects of M depletion on the three inflammogens); and 2)
the precaution of provoking the inflammatory response 24 h after
the last injection of the liposome preparation was taken. At variance
from MC depletion, a distinction between zymosan and LPS peritonitis
was observed following M depletion, such that LPS-induced PMN influx
was almost abolished (-90%), whereas the response to zymosan was
unexpectedly augmented. The former data are not surprising (19), but
the data obtained with zymosan prompted a series of experiments that is
discussed below. No modulation by either MCs or M of the TG response
was seen; for this reason this inflammogen was not used in subsequent
experiments. Overall, these observations confirm the important role
that resident MCs play in the initiation of the cellular component
characteristic of acute inflammation (7, 8, 10).
To extend these studies, we then investigated whether chemokine
generation could also be modulated by MCs in vivo as well as by PMN
influx. We chose to monitor the archetypal chemokines KC and mMCP-1
because both have been associated with PMN influx in the mouse (5, 17, 20). We found that in the zymosan peritonitis model resident MCs
accounted for a large part of the release of these two chemokines. The
reduction seen at the protein level after MC depletion is possibly
secondary to a repression at the gene level, although due to the
qualitative rather than quantitative value of the RT-PCR analysis, this
conclusion needs to be corroborated by other studies. The possibility
that MCs produce some soluble mediators that, in turn, cause KC and
mMCP-1 release from another cell type cannot be excluded. To our
knowledge this is the first study that shows evidence for a crucial
role of MCs in chemokine release during experimental inflammation in
vivo. To date, few in vitro studies have demonstrated the ability of
MCs to produce chemokines. In particular, human skin MCs and a human MC
line synthesize de novo IL-8, which is then stored in granules and
released upon activation (21). Similarly, an anti-IgE-dependent
secretion of MIP-1 from human MCs has been recently observed (22).
Finally, expression of multiple chemokine genes, including mMCP-1 and
IL-8, has been demonstrated again in a human MC leukemia cell line
(23).
In contrast to zymosan, i.p. injection of LPS led to the production of
KC and mMCP-1, which were not derived from M or MCs. It is very
likely that another cell type(s), such as endothelial cells (24) or
mesothelial cells (25, 26), could be responsible for the effect of LPS.
Mesothelial cells may be the best candidate, since they have been
recently implicated in the production of mMCP-1 in related models of
inflammation (25) and can also be present in small numbers in the
peritoneal cell pellet (where we could detect LPS-induced KC and mMCP-1
mRNA by RT-PCR analysis). In this particular condition, we have also
measured TNF- levels, but again LPS-induced release of this cytokine
was not altered by M depletion. Interestingly, the large majority of
TNF- produced in murine models of bacteria peritonitis derives from
peritoneal MCs rather than M (27).
The potentiating effect seen in M-depleted animals was even more
evident when a lower dose of zymosan was used, with an approximately
twofold increase in PMN extravasation at the 4 h point. It is very
likely that the cellular accumulation obtained with 1 mg of zymosan is
almost maximal for a single mouse, and it could only be potentiated by
40–50% in M-depleted animals. In contrast to the results with 1 mg
of zymosan and discussed above, the release of KC measured with the
lower dose of zymosan was almost entirely due to the resident M.
Together, these data indicate the existence of at least one endogenous
mediator released from the M that acts to down-regulate PMN
accumulation. IL-10 has been shown to reduce PMN elicitation in
experimental inflammation (28) and to inhibit the production of
proinflammatory cytokines and chemokines (29, 30). In the present study
immunoreactive IL-10 was detected in the zymosan exudates, and more
importantly, its levels were modulated by M, but not MC, depletion.
IL-10 KO mice have become available to help unravel the numerous
biological activities of this cytokine (31, 32). Disruption of the
IL-10 gene mimicked the effect of M removal in IL-10 wt mice in
terms of potentiation of PMN influx. A drastic reduction (>85%) in
peritoneal M numbers in IL-10 KO mice resulted in a further increase
in 4 h PMN influx, suggesting the involvement of M product(s),
other than IL-10, in these experimental conditions. A marked increase
in mMCP-1 and KC levels was also measured in IL-10 KO mice, but this
was not further modulated by M depletion, indicating different
mechanisms for control of leukocyte infiltration and chemokine
production. This hypothesis seems plausible, since twice as much mMCP-1
protein is found at the 8 h point in IL-10 KO animals.
In conclusion, this study compared three different inflammatory stimuli
in terms of 1) the profile of PMN accumulation in the mouse peritoneal
cavity, 2) chemokine generation, and 3) role of resident M and MCs.
We conclude that resident peritoneal MCs play a central role in the
production of the CXC chemokine KC and the CC chemokine mMCP-1 in the
acute inflammatory response induced by i.p. injection of zymosan. In
addition, endogenous IL-10 released from resident M plays a negative
modulatory role, with an effect on PMN accumulation and chemokine
generation. The existence of a stimulus and dose specificity for the
cell type(s) responsible for the rapid production of chemokines during
experimental inflammation is also proposed.
|
Footnotes |
|---|
2 Address correspondence and reprint requests to Dr. Mauro Perretti, Department of Biochemical Pharmacology, The William Harvey Research Institute, Charterhouse Square, London, U.K. EC1 M6BQ. E-mail address:
3 Abbreviations used in this paper: MCP, monocyte chemoattractant protein; MIP, macrophage inflammatory protein; m, murine; PMN, polymorphonuclear leukocyte; M, macrophage; MC, mast cell; wt, wild type; KO, knockout; TG, thioglycolate; Cl2MDP, dichloromethylene-bisphosphonate.
Received for publication June 3, 1998. Accepted for publication October 16, 1998.
|
References |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
. J. Immunol. 140:3144.[Abstract]
. J. Immunol. 159:3595.[Abstract]
by human mast cells: increased anti-IgE dependent secretion after IgE-dependent enhancement of mast cell IgE-binding activity. Lab. Invest. 77:185.[Medline]
. Nature 381:77.[Medline]
This article has been cited by other articles:
|
|
 |
|
  E. Kolaczkowska, W. Grzybek, N. van Rooijen, H. Piccard, B. Plytycz, B. Arnold, and G. Opdenakker Neutrophil elastase activity compensates for a genetic lack of matrix metalloproteinase-9 (MMP-9) in leukocyte infiltration in a model of experimental peritonitis J. Leukoc. Biol., March 1, 2009; 85(3): 374 - 381. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 C. K. Hu, F. Venet, D. S. Heffernan, Y. L. Wang, B. Horner, X. Huang, C.-S. Chung, S. H. Gregory, and A. Ayala The Role of Hepatic Invariant NKT Cells in Systemic/Local Inflammation and Mortality during Polymicrobial Septic Shock J. Immunol., February 15, 2009; 182(4): 2467 - 2475. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. J. Paul-Clark, S. K. McMaster, R. Sorrentino, S. Sriskandan, L. K. Bailey, L. Moreno, B. Ryffel, V. F. Quesniaux, and J. A. Mitchell Toll-like Receptor 2 Is Essential for the Sensing of Oxidants during Inflammation Am. J. Respir. Crit. Care Med., February 15, 2009; 179(4): 299 - 306. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. Leendertse, R. J.L. Willems, I. A.J. Giebelen, J. J.T.H. Roelofs, N. van Rooijen, M. J.M. Bonten, and T. van der Poll Peritoneal macrophages are important for the early containment of Enterococcus faecium peritonitis in mice Innate Immunity, February 1, 2009; 15(1): 3 - 12. [Abstract] [PDF]
|
|
|
|
 |
|
 X. Qiu, L. Zhu, and J. W. Pollard Colony-Stimulating Factor-1-Dependent Macrophage Functions Regulate the Maternal Decidua Immune Responses against Listeria monocytogenes Infections during Early Gestation in Mice Infect. Immun., January 1, 2009; 77(1): 85 - 97. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 R. A. Dean, J. H. Cox, C. L. Bellac, A. Doucet, A. E. Starr, and C. M. Overall Macrophage-specific metalloelastase (MMP-12) truncates and inactivates ELR+ CXC chemokines and generates CCL2, -7, -8, and -13 antagonists: potential role of the macrophage in terminating polymorphonuclear leukocyte influx Blood, October 15, 2008; 112(8): 3455 - 3464. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 R. Rajakariar, M. Hilliard, T. Lawrence, S. Trivedi, P. Colville-Nash, G. Bellingan, D. Fitzgerald, M. M. Yaqoob, and D. W. Gilroy From the Cover: Hematopoietic prostaglandin D2 synthase controls the onset and resolution of acute inflammation through PGD2 and 15-deoxy{Delta}12-14 PGJ2 PNAS, December 26, 2007; 104(52): 20979 - 20984. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. M. Burke, L. M. Ganley-Leal, A. Khatri, and L. M. Wetzler Neisseria meningitidis PorB, a TLR2 Ligand, Induces an Antigen-Specific Eosinophil Recall Response: Potential Adjuvant for Helminth Vaccines? J. Immunol., September 1, 2007; 179(5): 3222 - 3230. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. A Mitchell, M. J Paul-Clark, G. W Clarke, S. K McMaster, and N. Cartwright Critical role of toll-like receptors and nucleotide oligomerisation domain in the regulation of health and disease J. Endocrinol., June 1, 2007; 193(3): 323 - 330. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 D. N. Tziakas, G. K. Chalikias, C. O. Antonoglou, S. Veletza, I. K. Tentes, A. X. Kortsaris, D. I. Hatseras, and J. C. Kaski Apolipoprotein E Genotype and Circulating Interleukin-10 Levels in Patients With Stable and Unstable Coronary Artery Disease J. Am. Coll. Cardiol., December 19, 2006; 48(12): 2471 - 2481. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. C. Mullaly and P. Kubes The Role of TLR2 In Vivo following Challenge with Staphylococcus aureus and Prototypic Ligands J. Immunol., December 1, 2006; 177(11): 8154 - 8163. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
E. Kolaczkowska, A. Scislowska-Czarnecka, M. Chadzinska, B. Plytycz, N. van Rooijen, G. Opdenakker, and B. Arnold Enhanced early vascular permeability in gelatinase B (MMP-9)-deficient mice: putative contribution of COX-1-derived PGE2 of macrophage origin J. Leukoc. Biol., July 1, 2006; 80(1): 125 - 132. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. S. Damazo, S. Yona, R. J. Flower, M. Perretti, and S. M. Oliani Spatial and Temporal Profiles for Anti-Inflammatory Gene Expression in Leukocytes during a Resolving Model of Peritonitis J. Immunol., April 1, 2006; 176(7): 4410 - 4418. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. Nakayama, N. Mutsuga, L. Yao, and G. Tosato Prostaglandin E2 promotes degranulation-independent release of MCP-1 from mast cells J. Leukoc. Biol., January 1, 2006; 79(1): 95 - 104. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
B. E. Chatterjee, S. Yona, G. Rosignoli, R. E. Young, S. Nourshargh, R. J. Flower, and M. Perretti Annexin 1-deficient neutrophils exhibit enhanced transmigration in vivo and increased responsiveness in vitro J. Leukoc. Biol., September 1, 2005; 78(3): 639 - 646. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Matsukawa, S. Kudo, T. Maeda, K. Numata, H. Watanabe, K. Takeda, S. Akira, and T. Ito Stat3 in Resident Macrophages as a Repressor Protein of Inflammatory Response J. Immunol., September 1, 2005; 175(5): 3354 - 3359. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. W. Nasser, R. J. Marjoram, S. L. Brown, and R. M. Richardson Cross-Desensitization among CXCR1, CXCR2, and CCR5: Role of Protein Kinase C-{epsilon} J. Immunol., June 1, 2005; 174(11): 6927 - 6933. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. F. Cailhier, M. Partolina, S. Vuthoori, S. Wu, K. Ko, S. Watson, J. Savill, J. Hughes, and R. A. Lang Conditional Macrophage Ablation Demonstrates That Resident Macrophages Initiate Acute Peritoneal Inflammation J. Immunol., February 15, 2005; 174(4): 2336 - 2342. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 T.A. Silva, V.S. Lara, J.S. Silva, S.H.P. Oliveira, W.T. Butler, and F.Q. Cunha Macrophages and Mast Cells Control the Neutrophil Migration Induced by Dentin Proteins Journal of Dental Research, January 1, 2005; 84(1): 79 - 83. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
F. Bossi, F. Fischetti, V. Pellis, R. Bulla, E. Ferrero, T. E. Mollnes, D. Regoli, and F. Tedesco Platelet-Activating Factor and Kinin-Dependent Vascular Leakage as a Novel Functional Activity of the Soluble Terminal Complement Complex J. Immunol., December 1, 2004; 173(11): 6921 - 6927. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. P. Matzer, F. Rodel, R. M. Strieter, M. Rollinghoff, and H. U. Beuscher Constitutive expression of CXCL2/MIP-2 is restricted to a Gr-1high, CD11b+, CD62Lhigh subset of bone marrow derived granulocytes Int. Immunol., November 1, 2004; 16(11): 1675 - 1683. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T.-B. Kang, T. Ben-Moshe, E. E. Varfolomeev, Y. Pewzner-Jung, N. Yogev, A. Jurewicz, A. Waisman, O. Brenner, R. Haffner, E. Gustafsson, et al. Caspase-8 Serves Both Apoptotic and Nonapoptotic Roles J. Immunol., September 1, 2004; 173(5): 2976 - 2984. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. L. Lomas-Neira, C.-S. Chung, P. S. Grutkoski, E. J. Miller, and A. Ayala CXCR2 inhibition suppresses hemorrhage-induced priming for acute lung injury in mice J. Leukoc. Biol., July 1, 2004; 76(1): 58 - 64. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Stefanidakis, T. Ruohtula, N. Borregaard, C. G. Gahmberg, and E. Koivunen Intracellular and Cell Surface Localization of a Complex between {alpha}M{beta}2 Integrin and Promatrix Metalloproteinase-9 Progelatinase in Neutrophils J. Immunol., June 1, 2004; 172(11): 7060 - 7068. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. Takeshita, T. Yamasaki, S. Akira, F. Gantner, and K. B. Bacon Essential role of MHC II-independent CD4+ T cells, IL-4 and STAT6 in contact hypersensitivity induced by fluorescein isothiocyanate in the mouse Int. Immunol., May 1, 2004; 16(5): 685 - 695. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 R. L. Thurmond, P. J. Desai, P. J. Dunford, W.-P. Fung-Leung, C. L. Hofstra, W. Jiang, S. Nguyen, J. P. Riley, S. Sun, K. N. Williams, et al. A Potent and Selective Histamine H4 Receptor Antagonist with Anti-Inflammatory Properties J. Pharmacol. Exp. Ther., April 1, 2004; 309(1): 404 - 413. [Abstract] [Full Text]
|
|
|
|
|
|
B. T. Edelson, Z. Li, L. K. Pappan, and M. M. Zutter Mast cell-mediated inflammatory responses require the {alpha}2{beta}1 integrin Blood, March 15, 2004; 103(6): 2214 - 2220. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. Di Filippo, F. Rossi, S. Rossi, and M. D'Amico Cannabinoid CB2 receptor activation reduces mouse myocardial ischemia-reperfusion injury: involvement of cytokine/chemokines and PMN J. Leukoc. Biol., March 1, 2004; 75(3): 453 - 459. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. A. Willment, H.-H. Lin, D. M. Reid, P. R. Taylor, D. L. Williams, S. Y. C. Wong, S. Gordon, and G. D. Brown Dectin-1 Expression and Function Are Enhanced on Alternatively Activated and GM-CSF-Treated Macrophages and Are Negatively Regulated by IL-10, Dexamethasone, and Lipopolysaccharide J. Immunol., November 1, 2003; 171(9): 4569 - 4573. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. Murata, M. Inami, A. Hasegawa, S. Kubo, M. Kimura, M. Yamashita, H. Hosokawa, T. Nagao, K. Suzuki, K. Hashimoto, et al. CD69-null mice protected from arthritis induced with anti-type II collagen antibodies Int. Immunol., August 1, 2003; 15(8): 987 - 992. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Hadjur and F. R. Jirik Increased sensitivity of Fancc-deficient hematopoietic cells to nitric oxide and evidence that this species mediates growth inhibition by cytokines Blood, May 15, 2003; 101(10): 3877 - 3884. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. A. Garhart, F. P. Heinzel, S. J. Czinn, and J. G. Nedrud Vaccine-Induced Reduction of Helicobacter pylori Colonization in Mice Is Interleukin-12 Dependent but Gamma Interferon and Inducible Nitric Oxide Synthase Independent Infect. Immun., February 1, 2003; 71(2): 910 - 921. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. Schramm, T. Schaefer, M. D. Menger, and H. Thorlacius Acute mast cell-dependent neutrophil recruitment in the skin is mediated by KC and LFA-1: inhibitory mechanisms of dexamethasone J. Leukoc. Biol., December 1, 2002; 72(6): 1122 - 1132. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. S. HUBER-LANG, N. C. RIEDEMAN, J. V. SARMA, E. M. YOUNKIN, S. R. McGUIRE, I. J. LAUDES, K. T. LU, R.-F. GUO, T. A. NEFF, V. A. PADGAONKAR, et al. Protection of innate immunity by C5aR antagonist in septic mice FASEB J, October 1, 2002; 16(12): 1567 - 1574. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. Kurosaka, N. Watanabe, and Y. Kobayashi Potentiation by human serum of anti-inflammatory cytokine production by human macrophages in response to apoptotic cells J. Leukoc. Biol., June 1, 2002; 71(6): 950 - 956. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Dobrina, M. Pausa, F. Fischetti, R. Bulla, E. Vecile, E. Ferrero, A. Mantovani, and F. Tedesco Cytolytically inactive terminal complement complex causes transendothelial migration of polymorphonuclear leukocytes in vitro and in vivo Blood, January 1, 2002; 99(1): 185 - 192. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 C. de Coupade, M. N. Ajuebor, F. Russo-Marie, M. Perretti, and E. Solito Cytokine Modulation of Liver Annexin 1 Expression during Experimental Endotoxemia Am. J. Pathol., October 1, 2001; 159(4): 1435 - 1443. [Abstract] [Full Text]
|
|
|
|
|
|
M. Delgado and D. Ganea Inhibition of Endotoxin-Induced Macrophage Chemokine Production by Vasoactive Intestinal Peptide and Pituitary Adenylate Cyclase-Activating Polypeptide In Vitro and In Vivo J. Immunol., July 15, 2001; 167(2): 966 - 975. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Deckert, S. Soltek, G. Geginat, S. Lutjen, M. Montesinos-Rongen, H. Hof, and D. Schluter Endogenous Interleukin-10 Is Required for Prevention of a Hyperinflammatory Intracerebral Immune Response in Listeria monocytogenes Meningoencephalitis Infect. Immun., July 1, 2001; 69(7): 4561 - 4571. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
I. Guleria and J. W. Pollard Aberrant Macrophage and Neutrophil Population Dynamics and Impaired Th1 Response to Listeria monocytogenes in Colony-Stimulating Factor 1-Deficient Mice Infect. Immun., March 1, 2001; 69(3): 1795 - 1807. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. Ikeda, H. Saito, K. Fukatsu, T. Inoue, I. Han, S. Furukawa, T. Matsuda, and A. Hidemura Dietary Restriction Impairs Neutrophil Exudation by Reducing CD11b/CD18 Expression and Chemokine Production Arch Surg, March 1, 2001; 136(3): 297 - 304. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
E. Kolaczkowska, R. Seljelid, and B. Plytycz Role of mast cells in zymosan-induced peritoneal inflammation in Balb/c and mast cell-deficient WBB6F1 mice J. Leukoc. Biol., January 1, 2001; 69(1): 33 - 42. [Abstract] [Full Text]
|
|
|
|
|
|
J. Witowski, K. Pawlaczyk, A. Breborowicz, A. Scheuren, M. Kuzlan-Pawlaczyk, J. Wisniewska, A. Polubinska, H. Friess, G. M. Gahl, U. Frei, et al. IL-17 Stimulates Intraperitoneal Neutrophil Infiltration Through the Release of GRO{alpha} Chemokine from Mesothelial Cells J. Immunol., November 15, 2000; 165(10): 5814 - 5821. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 R. A. Terkeltaub IL-10: An "Immunologic Scalpel" for Atherosclerosis? Arterioscler. Thromb. Vasc. Biol., December 1, 1999; 19(12): 2823 - 2825. [Full Text] [PDF]
|
|
|
|
|
|
N. Vergnolle Proteinase-Activated Receptor-2-Activating Peptides Induce Leukocyte Rolling, Adhesion, and Extravasation In Vivo J. Immunol., November 1, 1999; 163(9): 5064 - 5069. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. Warny, S. Aboudola, S. C. Robson, J. Sevigny, D. Communi, S. P. Soltoff, and C. P. Kelly P2Y6 Nucleotide Receptor Mediates Monocyte Interleukin-8 Production in Response to UDP or Lipopolysaccharide J. Biol. Chem., July 6, 2001; 276(28): 26051 - 26056. [Abstract] [Full Text] [PDF]
|
|
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
