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in Mouse Macrophages1
*
Department of Medicine, Johns Hopkins University School of Medicine, Baltimore, MD 21205; and
Department of Medicine, University of Michigan Medical Center, Ann Arbor, MI 48109
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Abstract |
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 Top Abstract IntroductionMaterials and MethodsResultsDiscussionReferences |
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on
HA-induced chemokine gene expression in primary mouse macrophages. We
found that IL-10 and IFN- independently inhibit HA-induced
expression of macrophage inflammatory protein-1
(MIP-1),
MIP-1ß, and KC at both the mRNA and protein levels. Whereas IL-10
inhibited most of the HA-induced chemokines tested, IFN- selectively
inhibited only MIP-1, MIP-1ß, and KC. This inhibition did not
require prestimulation and occurred even when the cytokines were added
up to 3 h after stimulation with HA. For MIP-1, the inhibition
by IFN- occurred at the level of transcription, whereas IL-10
predominantly decreased the stability of MIP-1 mRNA. IFN- and
IL-10 equally inhibited macrophage expression of MIP-1ß mRNA at the
level of transcription, but MIP-1ß mRNA stability was decreased to a
greater extent by IL-10. These data identify a previously unrecognized
role for IL-10 and IFN- as regulators of ECM-induced macrophage
expression of inflammatory chemokines. |
Introduction |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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Recent studies suggest a role for ECM components in the activation of inflammatory macrophages (2). In chronic inflammation there is increased production and degradation of the ECM (3). ECM fragments have been shown to have different biologic activities than those of their larger, native precursors (4, 5, 6). Fragments of the ECM components collagen and fibronectin have been shown to have proinflammatory properties (7). Thus, ECM fragments generated at sites of inflammation may play a role in macrophage activation (2). Recent studies have shown that low m.w. fragments of the ECM component HA are capable of inducing the expression of a number of inflammatory gene products, including cytokines, chemokines, reactive nitrogen species, and growth factors (8, 9, 10).
HA is a nonsulfated glycosaminoglycan polymer made up of repeating
disaccharide units of (ß,1
4)-D-glucuronic acid-(ß,
13)-N-acetyl-D-glucosamine. HA is a
ubiquitously distributed component of the ECM and in its native form
exists as a high m.w. polymer (11, 12). In normal tissues this high
m.w. HA functions in water homeostasis, plasma protein distribution,
and matrix structuring (11). At sites of inflammation and tissue injury
there is an accumulation of lower m.w. HA species (13, 14, 15). Recent
studies have suggested that these lower m.w. forms of HA may stimulate
macrophages recruited to sites of inflammation to produce important
mediators of tissue injury and repair (8). The excessive production of
inflammatory mediators by activated macrophages could result in the
perpetuation of a chronic inflammatory state culminating in tissue
fibrosis. A recently identified and rapidly enlarging group of
inflammatory mediators that have been shown to play an important role
in chronic inflammatory states are the chemokines (16). MIP-1 and
MIP-1ß are two members of the C-C class of chemokines that have been
directly implicated in the pathogenesis of chronic inflammation in
joint and lung (17). Recent work from our laboratory has suggested that
one mechanism for inducing the expression of MIP-1 and MIP-1ß at
sites of inflammation may be through the effect of low m.w.
HA on inflammatory macrophages (8). The mechanisms that regulate
ECM-induced inflammatory gene expression in macrophages are
unknown.
Two cytokines that are known modulators of macrophage effector
functions and regulators of the inflammatory response are IL-10 and
IFN- (18, 19, 20). IL-10 has been shown to deactivate macrophages by
inhibiting production of proinflammatory cytokines, such as TNF-
(18, 20, 21, 22), and reactive oxygen intermediates (22). IFN- enhances
certain macrophage functions, such as microbiocidal and tumoricidal
activity, through the production of reactive oxygen intermediates and
reactive nitrogen intermediates (23, 24). Despite its many
proinflammatory roles, IFN- also inhibits the expression of certain
LPS-induced chemokines, such as MCP-1 and KC in macrophages (25). In
further support of its selective anti-inflammatory properties,
IFN- has been shown to ameliorate bleomycin-induced lung
inflammation and fibrosis in animal models (26).
The purpose of this investigation was to determine the effects of IL-10
and IFN- on chemokine gene expression induced by low m.w. HA
fragments. Our results show that IL-10 and IFN- inhibit HA-induced
chemokine gene expression in primary murine macrophages. IL-10 was
found to inhibit most HA-induced chemokines, while IFN- was
selective and inhibited only MIP-1, MIP-1ß, and KC. Furthermore,
IL-10 and IFN- appear to inhibit chemokine gene expression by
altering both mRNA stability and gene transcription.
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Materials and Methods |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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Mouse BMDM were isolated, as previously described (9), from female C3H/HeJ LPS-hyporesponsive mice purchased from The Jackson Laboratory (Bar Harbor, ME). After harvest, cells (11 x 106 cells/dish) were cultured for 5 days in DMEM supplemented with 10% heat-inactivated, low LPS FBS; 15% L cell medium; and 1% penicillin-streptomycin/1% glutamine (Biofluids, Rockville, MD) at 37°C under 8% CO2. Thioglycolate-elicited peritoneal macrophages were isolated from female C3H/HeJ mice 4 days after injection of 2 ml of sterile thioglycolate (Sigma Chemical Co., St. Louis, MO). The cells were allowed to adhere overnight in RPMI 1640 supplemented with 10% heat-inactivated, low LPS FBS and 1% penicillin-streptomycin/1% glutamine before use. To exclude the effects of contaminating LPS on experimental conditions, cell stimulation was conducted in the presence of polymyxin B (10 µg/ml; Calbiochem Novabiochem, La Jolla, CA). All experiments were conducted in serum-free RPMI 1640 with 1% penicillin-streptomycin/1% glutamine, so as to minimize the effects of serum stimulation (27).
Chemicals and reagents
Purified HA fragments from human umbilical cords were purchased
from ICN Biomedicals, Inc. (Costa Mesa, CA). The HA-ICN preparation is
free of protein (<2%) and free of chondroitin sulfate (<3%), and we
have previously determined its peak molecular size to be approximately
200,000 Da (28). Recombinant mouse IFN- (sp. act., 3.0 x
105 U/ml; endotoxin level, <0.2 ng/mg) was obtained
from Genzyme Corp. (Cambridge, MA), recombinant mouse IL-10 (5 ng/ml)
was purchased from R&D Systems (Minneapolis, MN), and actinomycin D (50
µg/ml) and cycloheximide (10 µg/ml) were obtained from Sigma
Chemical Co. Polymyxin B was purchased from Calbiochem Novabiochem.
Stock solutions of reagents were tested for LPS contamination using the
Limulus amoebocyte assay (Sigma).
Northern analysis of mRNA production
RNA was extracted from confluent cell monolayers using 4 M guanidine isothiocyanate and was purified by centrifugation through 5.7 M cesium chloride for 12 to 18 h at 35,000 rpm as previously described (9). Ten micrograms of total RNA was electrophoresed under denaturing conditions through a 1% formaldehyde-containing agarose gel, and RNA was transferred to Nytran (Schleicher and Schuell, Keene, NH) hybridization filters. Blots were briefly rinsed in 5x SSC, RNA was cross-linked to the filter by UV cross-linking (Stratagene, La Jolla, CA), and blots were hybridized overnight with 106 cpm/ml of 32P-labeled DNA labeled by the random prime method (Amersham, Arlington Heights, IL). Following hybridization, blots were washed once in 2x SSC/0.1% SDS at room temperature for 30 min with shaking, then washed twice in 0.1x SSC/0.1% SDS at 50°C with shaking for 20 min each wash. Blots were exposed at -70°C against Kodak XAR diagnostic film. Differences in RNA loading were documented by hybridizing selected blots with 32P-labeled cDNA for aldolase (29). Densitometric scanning was performed using a Molecular Dynamics Personal Densitometer SI (Sunnyvale, CA).
Determination of chemokine protein production
BMDM were plated at a density of 11 x
106/ml, and following stimulation with HA and/or IL-10
and/or IFN-, supernatants were collected at given time points. ELISA
measurements were performed as previously described (30).
Nuclear run-on
Nuclei from confluent monolayers of elicited peritoneal macrophages were harvested by scraping in ice-cold PBS and subsequently isolated by centrifugation through a sucrose cushion (31). Nuclei were then incubated for 30 min with 1 M DTT, 20 mM NTP, and 100 µCi of [32P]UTP in transcription buffer. The nuclei received a cold UTP chase for 10 min before the reaction was stopped by addition of termination buffer, DNase (Promega, Madison, WI), and RNase inhibitor (Boehringer Mannheim Corp., Indianapolis, IN). The nuclei were then incubated with transfer RNA (Sigma Chemical Co.) for 15 min before addition of 10% SDS, 0.2 M EDTA, and proteinase K (Sigma). After 15-min incubation, the RNA was extracted with phenol/chloroform/isoamyl alcohol, precipitated with 20% TCA, washed with 5% TCA/5% inorganic pyrophosphate, dissolved in 0.1% N-tris (hydroxymethyl)methyl-2-aminoethanesulfonic acid (TES), and precipitated for a second time with 4 M sodium acetate and 100% ethanol at -80°C for 30 min. Purified radiolabeled RNA was washed once in 70% ethanol and dried with a Speed-Vac concentrator (Savant Instruments, Farmingdale, NY), and resuspended in 100 µl of diethylpyrocarbonate-treated water. Five microliters of the radioactive RNA was counted, and all samples were normalized for counts using hybridization fluid. Normalized sampled were hybridized with prehybridized Optitran-S membranes (Schleicher and Schuell) containing the cDNAs of interest. Blots were hybridized for 3 to 4 days and then washed once in 2x SSC/0.1% SDS at room temperature for 5 min with shaking and washed twice in 0.1x SSC/0.1% SDS at 50°C with shaking (20 min each wash). The blots were exposed and quantitated with a PhosphorImager (Molecular Dynamics, Sunnyvale, CA).
Statistical analysis
Statistical analysis was preformed using the StatView analysis of variance program (Abacus Concepts, Inc., Berkeley, CA).
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Results |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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inhibit HA-induced MIP-1 and MIP-1ß mRNA
expression in primary mouse macrophages
We investigated the effects of IL-10 and IFN- on
HA-induced chemokine mRNA expression by BMDM and inflammatory
thioglycolate-elicited peritoneal macrophages from
C3H/HeJ LPS hyporesponsive mice. We have previously
shown that HA fragments (200,000 Da) induce the expression of a variety
of inflammatory mediators, including MIP-1, MIP-1ß, IP-10, IL-12,
RANTES, MCP-1, TNF-, KC, and nitric oxide in an alveolar macrophage
cell line (8). To assess the effects of IL-10 and IFN- on HA-induced
inflammatory gene expression, BMDM- and thioglycolate-elicited
peritoneal macrophages were simultaneously stimulated with HA in the
presence or the absence of cytokine for 6 h, mRNA was isolated,
and Northern analysis was performed. As shown in Figure 1
, IL-10 inhibited the HA-induced
expression of MIP-1, MIP-1ß, and TNF- in both BMDM (Fig. 1a) and thioglycolate-elicited peritoneal macrophages
(Fig. 1b). However, IFN- inhibited HA-induced mRNA
production of only MIP-1 and MIP-1ß, while enhancing RANTES and
TNF- expression in both BMDM- and thioglycolate-elicited peritoneal
macrophages. Both IL-10 and IFN- independently inhibited HA-induced
KC mRNA in thioglycolate-elicited peritoneal macrophages. The
expression of HA-induced MCP-1 and RANTES mRNA by these macrophages was
minimally effected by IL-10, but appeared to be slightly enhanced by
IFN-. When added together, IL-10 and IFN- exhibited a complete
inhibition of HA-induced MIP-1 and MIP-1ß mRNA expression in
BMDM-elicited peritoneal macrophages and of HA-induced MIP-1 and KC
in thioglycolate-elicited peritoneal macrophages, suggesting that the
cytokines cause inhibition by distinct mechanisms. These data suggest
that IL-10 and IFN- are negative regulators of HA-induced chemokine
expression, with IFN- targeting a select group (MIP-1, MIP-1ß,
and KC).
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exhibit a time-dependent and bimodal effect on
HA-induced expression of MIP-1 and MIP-1ß mRNA
To further delineate the effects of IL-10 and IFN- on
HA-induced expression of MIP-1 and MIP-1ß, we stimulated BMDM
simultaneously with HA in the presence or the absence of cytokine for
varying time intervals. As shown in Figure 2, an unexpected bimodal effect of IL-10
and IFN- on HA-induced expression of MIP-1 and MIP-1ß mRNA was
observed. Early on and peaking at 1 h, IL-10 and IFN- enhanced
HA-induced mRNA expression of these chemokines. No effect was observed
with the cytokines alone (data not shown). However, after 3 h,
IL-10 and IFN- began to inhibit HA-induced mRNA expression of
MIP-1 (Fig. 2), whereas they did not begin to inhibit MIP-1ß
expression until 4 h after stimulation (data not shown). The
inhibition of both MIP-1 and MIP-1ß by IL-10 and IFN- was
maximal after 6 h of stimulation. These results suggest that there
is a differential response in chemokine steady state mRNA levels of
HA-stimulated BMDM in the presence of IL-10 and IFN-. Identical
findings were observed for thioglycolate-elicited peritoneal
macrophages (data not shown).
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inhibit HA-induced MIP-1 and MIP-1ß protein
production in primary mouse macrophages
Having identified a bimodal effect of IL-10 and IFN- on
HA-induced chemokine mRNA, we investigated chemokine production at the
protein level. BMDM were simultaneously stimulated with HA in the
presence or the absence of cytokine for varying time intervals.
Chemokine levels were determined in the supernatants by ELISA. Figure 3 shows that HA-induced MIP-1 (Fig. 3a) and MIP-1ß (Fig. 3b)
chemokine production was maximal at 6 to 9 h and was markedly
inhibited in the presence of IL-10, IFN-, or both at 9 h (Fig. 3c). Thus, despite the early stimulatory effect of
IL-10 and IFN- on HA-induced mRNA expression of MIP-1 and
MIP-1ß, only an inhibitory effect was observed at the protein level.
Identical results were obtained for inflammatory peritoneal
macrophages, except the secreted levels of MIP-1 and MIP-1ß
protein were greater (data not shown).
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required to
inhibit HA-induced chemokine gene expression. As shown in Figure 4, the inhibition of HA-induced chemokine
gene expression was seen with as little as 1 ng/ml IL-10 and 10 U/ml
IFN- and was maximal at 5 ng/ml IL-10 and 100 U/ml IFN-. Thus,
low concentrations of these regulatory cytokines were able to evoke
maximal inhibition of HA-induced MIP-1 and MIP-1ß gene expression
in primary macrophages.
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inhibition of HA-induced MIP-1 and MIP-1ß gene expression
To further dissect the mechanism for the observed inhibition, we
examined the relationship between time of exposure of cytokine and
inhibition of HA-induced chemokine gene expression. We preincubated
BMDM in the presence of IL-10 and IFN- for 1, 2, or 3 h before
the addition of HA fragments. The cells were then allowed to incubate
with all stimuli for 6 h. Figure 5
shows that prestimulation was not necessary for the inhibition of
HA-induced MIP-1 and MIP-1ß mRNA expression by IL-10 and IFN-.
In fact, for IFN-, the longer the period of prestimulation, the less
inhibition of HA-induced gene expression was observed. These results
suggest that the effect of IL-10 and IFN- on HA-induced gene
expression occurs immediately upon stimulation, thus making the
requirement for new protein synthesis less likely.
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could
still inhibit HA-induced MIP-1 and MIP-1ß mRNA expression in BMDM
if given after the cells had already been stimulated with HA. BMDM were
stimulated with HA for 0, 1, and 3 h before addition of IL-10
and/or IFN-. RNA was isolated after 6-h total stimulation with HA.
Figure 6 shows that IL-10 and IFN-
still inhibited HA-induced gene expression of MIP-1 and MIP-1ß
even when added 3 h after the HA fragments. These results suggest
that the cytokine inhibition of HA-induced chemokine expression is
effective even after HA has initiated gene expression.
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and MIP-1ß by IL-10 and IFN-
To examine the role of new protein synthesis on the
inhibition of HA-induced mRNA expression of MIP-1 and MIP-1ß by
IL-10 and IFN-, we pretreated BMDM with CHX for 30 min before the
addition of HA and/or IL-10 and IFN- for 6 h. We found that, as
has been previously described (32), CHX itself markedly induced
MIP-1 mRNA expression making in impossible to determine whether new
protein synthesis is required for the cytokine inhibitory effect (data
not shown). MIP-1ß mRNA was not markedly increased by CHX alone, and
the inhibition by IL-10 and IFN- was slightly reduced (data not
shown). While failing to elucidate the role of new protein synthesis in
cytokine inhibition, the results do suggest that the HA induction of
MIP-1ß may be regulated differently than that of MIP-1.
We then sought to determine whether the mechanism of cytokine
inhibition of HA-stimulated chemokine gene expression was occurring at
the level of mRNA stability by performing experiments in the presence
of actinomycin D. Actinomycin D added before stimulation of BMDM with
HA completely blocked the induction of chemokine gene expression,
indicating that the effect of HA on chemokine mRNA expression is at the
level of transcription (data not shown). Elicited peritoneal
macrophages were stimulated with HA and/or IL-10/IFN- for 2 to
3 h before actinomycin D was added, and RNA was isolated at 1, 2,
and 4 h after addition of actinomycin D. As shown in Figure 7, IL-10 significantly shortened the
half-life of both HA-induced MIP-1 (Fig. 7a) and
MIP-1ß (Fig. 7b). IFN- also appears to have an
effect on the half-lives of these chemokine mRNAs, but to a lesser
extent than IL-10. These results indicate that the IL-10 inhibition of
HA-induced chemokine gene expression is predominantly at the level of
mRNA stability.
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on HA-induced MIP-1
and MIP-1ß gene expression in elicited peritoneal macrophages
To examine the direct effect of IL-10 and IFN- on HA-induced
chemokine gene transcription we performed nuclear run-on assays. We
radiolabeled the mRNA transcribed in nuclei isolated from elicited
peritoneal macrophages stimulated with HA, IL-10, and/or IFN- for
3 h. Figure 8a shows that
HA markedly induces transcription of both MIP-1 and MIP-1ß.
IFN- markedly inhibits HA-induced MIP-1 and MIP-1ß gene
transcription (Fig. 8b). IL-10 also exhibited an
effect on chemokine transcription equal to that of IFN- for
MIP-1ß, but to a lesser extent than that of IFN- on MIP-1 (Fig. 8b). These data are presented quantitatively in
Figure 8c.
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Discussion |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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as
regulators of ECM-induced chemokine gene expression. IL-10 and IFN-
independently inhibited the induction of the chemokines MIP-1,
MIP-1ß, and KC by low m.w. HA fragments. IL-10 inhibited nearly all
HA-induced chemokines tested, but IFN- selectively inhibited only a
subset of HA-induced chemokines. Thus, IL-10 appears to be a general
down-regulator of ECM-induced chemokine gene expression, whereas
IFN- can either inhibit or enhance specific HA-induced
chemokines.
Interestingly, both cytokines enhanced the HA-induced steady state mRNA
levels of MIP-1 and MIP-1ß at early time points. This increased
mRNA, however, does not appear to be translated into secreted protein.
The significance of this bimodal effect of IL-10 and IFN- on
HA-induced chemokine mRNA expression is unclear. The early enhancement
occurs only at the RNA level and may be the result of transient changes
in mRNA stability, since it is not reflected in the overall inhibitory
effect of IL-10 and IFN- on HA-induced expression of chemokine
protein secretion.
To further characterize the mechanisms by which IL-10 and IFN-
inhibited HA-induced chemokine gene expression in inflammatory
macrophages, we performed pre- and poststimulation experiments.
Prestimulation with IL-10 and/or IFN- diminished the peak inhibitory
effect of these cytokines on HA-induced chemokine steady state mRNA
levels. Furthermore, IL-10 and IFN- can effectively inhibit
HA-induced mRNA of MIP-1 and MIP-1ß even when given up to 3 h
after stimulation. Thus, maximal inhibition of these chemokines was
seen with the simultaneous administration of IL-10 and IFN- with HA.
These results suggest that IL-10 and IFN- have an immediate effect
on HA-induced chemokine mRNA expression and are most likely not working
through a secondary effector molecule.
The inhibitory effects of IL-10 and IFN- on MIP-1 and MIP-1ß
mRNA expression appear to be due to the differing effects the cytokines
have on mRNA stability and transcription. Both IL-10 and IFN-
shorten the half-lives of MIP-1 and MIP-1ß mRNA induced by HA. The
mRNA stability of both MIP-1 and MIP-1ß is significantly decreased
in the presence of IL-10 (70% reduction in half-life) and to a lesser
degree with IFN- (30% reduction). At the level of transcription,
these cytokines have slightly different effects on MIP-1 and
MIP-1ß gene expression. IFN- appears to exhibit a more significant
inhibition of HA-induced transcription of MIP-1 (60–70%) than
MIP-1ß (50%). In contrast, IL-10 caused a greater inhibition of
HA-induced MIP-1ß transcription (50%) than of that for MIP-1
(30–40%). In fact, HA-induced MIP-1ß transcription was equally
inhibited by both IFN- and IL-10 (50%). These results suggest that
the profound inhibition of steady state HA-induced MIP-1 and
MIP-1ß mRNA expression by IL-10 and IFN- is due to effects on both
mRNA stability and transcription.
IL-10 is a deactivator of numerous macrophage effector cell functions. In fact, IL-10 not only inhibits LPS-induced production of various chemokines by human macrophages (33), it also prevents lethal endotoxemia in mice (34) and immune complex-induced lung injury in rats (35). The mechanisms of IL-10-suppressive effects are incompletely understood. In this report we show that IL-10 has a general inhibitory effect on HA-induced chemokine expression, and that this inhibition, although in part occurring at the level of transcription, is predominantly at the level of decreased mRNA stability. These results are consistent with work by other investigators, who found that IL-10 enhanced the degradation of LPS-induced cytokine mRNA in macrophages (36).
Despite its many proinflammatory roles, IFN- also inhibits the
secretion of certain macrophage products. Oliveira et al. have shown
that IFN- inhibits TNF--induced IL-8 secretion from human
fibroblasts (37), and Ohmori et al. have shown that IFN- inhibits
the LPS-induced chemokines MCP-1 and KC by murine macrophages (25). In
addition, IFN- has also been shown to block the expression of HA-
and TNF--induced insulin-like growth factor I by BMDM (38). These
data suggest that IFN- may selectively down-regulate a subset of
proinflammatory genes, such as IL-8, MIP-1, and KC. Our results show
that IFN- has a differential effect on HA-induced chemokine gene
expression. IFN- inhibits certain HA-induced chemokines, such as
MIP-1, MIP-1ß, and KC. In the chronic inflammatory state produced
by bleomycin-induced lung injury in mice, previous investigators have
found that there is increased production of both the inflammatory
chemokines MIP-1/ß and KC and the low m.w. HA fragments (39, 40, 41).
Likewise, IFN- has been been shown to decrease lung fibrosis
following the instillation of bleomycin in the lungs of mice (26).
Thus, it is interesting to speculate that perhaps part of the
amelioration of bleomycin-induced lung fibrosis in mice may be from the
inhibitory effect of IFN- on HA-induced expression of these
profibrotic chemokines.
We have previously shown that the low m.w. HA fragments induce the
expression of inflammatory genes through the NF-
B/I-B
transcriptional regulatory system (8, 40, 42). Interestingly, the
inhibitory effects of both IL-10 and IFN- have been suggested to
occur by the inhibition of NF-B activation (37, 41). Recently, IL-10
has been shown to inhibit nuclear localization of NF-B in human
monocytes stimulated by LPS or TNF- (33). Similarly, recent
reports have shown that IFN- inhibits the expression of IL-8
from human fibroblasts and that this inhibition requires a NF-B
binding site in the IL-8 promoter (37, 41). Thus, IL-10 and IFN- may
in part be inhibiting HA-induced chemokine gene expression through
alterations in the NF-B/I-B transcriptional regulatory system.
This may be relevant in the case of MIP-1, which contains a proximal
NF-B site, but there is no NF-B site in the proximal MIP-1ß
promoter. This further supports the possibility of distinct mechanisms
regulating the expression of these two chemokines.
In conclusion, determining whether the marked inhibition of ECM-induced
MIP-1, MIP-1ß, and KC expression by IL-10 and IFN- observed in
vitro occurs in vivo and identifying the molecular mechanisms that
inhibit ECM-induced chemokine gene expression may offer new approaches
to controlling chronic tissue inflammation and fibrosis.
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Footnotes |
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2 Address correspondence and reprint requests to Dr. Paul W. Noble, Yale University School of Medicine, Veteran’s Administration Connecticut Healthcare System, Pulmonary Section/111A, 950 Campbell Avenue, West Haven, CT 06516. E-mail address:
3 Abbreviations used in this paper: ECM, extracellular matrix; HA, hyaluronan; MIP, macrophage inflammatory protein; MCP-1, monocyte chemoattractant protein-1; BMDM, bone marrow-derived macrophages; CHX, cycloheximide; NF-B, nuclear factor-B.
Received for publication September 3, 1997. Accepted for publication November 25, 1997.
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  P. L. Bollyky, B. A. Falk, R. P. Wu, J. H. Buckner, T. N. Wight, and G. T. Nepom Intact extracellular matrix and the maintenance of immune tolerance: high molecular weight hyaluronan promotes persistence of induced CD4+CD25+ regulatory T cells J. Leukoc. Biol., September 1, 2009; 86(3): 567 - 572. [Abstract] [Full Text] [PDF]
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 K. A. Scheibner, S. Boodoo, S. Collins, K. E. Black, Y. Chan-Li, P. Zarek, J. D. Powell, and M. R. Horton The Adenosine A2a Receptor Inhibits Matrix-Induced Inflammation in a Novel Fashion Am. J. Respir. Cell Mol. Biol., March 1, 2009; 40(3): 251 - 259. [Abstract] [Full Text] [PDF]
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T. L. Adair-Kirk, J. J. Atkinson, D. G. Kelley, R. H. Arch, J. H. Miner, and R. M. Senior A Chemotactic Peptide from Laminin {alpha}5 Functions as a Regulator of Inflammatory Immune Responses via TNF{alpha}-mediated Signaling J. Immunol., February 1, 2005; 174(3): 1621 - 1629. [Abstract] [Full Text] [PDF]
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T. S. Wilkinson, S. Potter-Perigo, C. Tsoi, L. C. Altman, and T. N. Wight Pro- and Anti-Inflammatory Factors Cooperate to Control Hyaluronan Synthesis in Lung Fibroblasts Am. J. Respir. Cell Mol. Biol., July 1, 2004; 31(1): 92 - 99. [Abstract] [Full Text] [PDF]
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 K. N. Sugahara, T. Murai, H. Nishinakamura, H. Kawashima, H. Saya, and M. Miyasaka Hyaluronan Oligosaccharides Induce CD44 Cleavage and Promote Cell Migration in CD44-expressing Tumor Cells J. Biol. Chem., August 22, 2003; 278(34): 32259 - 32265. [Abstract] [Full Text] [PDF]
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P. E. Pummill and P. L. DeAngelis Alteration of Polysaccharide Size Distribution of a Vertebrate Hyaluronan Synthase by Mutation J. Biol. Chem., May 23, 2003; 278(22): 19808 - 19814. [Abstract] [Full Text] [PDF]
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 G. M. Turino and J. O. Cantor Hyaluronan in Respiratory Injury and Repair Am. J. Respir. Crit. Care Med., May 1, 2003; 167(9): 1169 - 1175. [Full Text] [PDF]
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 M. Miura, T. El-Sawy, and R. L. Fairchild Neutrophils Mediate Parenchymal Tissue Necrosis and Accelerate the Rejection of Complete Major Histocompatibility Complex-Disparate Cardiac Allografts in the Absence of Interferon-{gamma} Am. J. Pathol., February 1, 2003; 162(2): 509 - 519. [Abstract] [Full Text] [PDF]
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M. R. Horton, S. Boodoo, and J. D. Powell NF-kappa B Activation Mediates the Cross-talk between Extracellular Matrix and Interferon-gamma (IFN-gamma ) Leading to Enhanced Monokine Induced by IFN-gamma (MIG) Expression in Macrophages J. Biol. Chem., November 8, 2002; 277(46): 43757 - 43762. [Abstract] [Full Text] [PDF]
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M. E. Mummert, D. Mummert, D. Edelbaum, F. Hui, H. Matsue, and A. Takashima Synthesis and Surface Expression of Hyaluronan by Dendritic Cells and Its Potential Role in Antigen Presentation J. Immunol., October 15, 2002; 169(8): 4322 - 4331. [Abstract] [Full Text] [PDF]
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T. D. Camenisch and J. A. McDonald Hyaluronan . Is Bigger Better? Am. J. Respir. Cell Mol. Biol., October 1, 2000; 23(4): 431 - 433. [Full Text]
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M. R. Horton, M. A. Olman, C. Bao, K. E. White, A. M. K. Choi, B.-Y. Chin, P. W. Noble, and C. J. Lowenstein Regulation of plasminogen activator inhibitor-1 and urokinase by hyaluronan fragments in mouse macrophages Am J Physiol Lung Cell Mol Physiol, October 1, 2000; 279(4): L707 - L715. [Abstract] [Full Text] [PDF]
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 M. E. Mummert, M. Mohamadzadeh, D. I. Mummert, N. Mizumoto, and A. Takashima Development of a Peptide Inhibitor of Hyaluronan-Mediated Leukocyte Trafficking J. Exp. Med., September 18, 2000; 192(6): 769 - 780. [Abstract] [Full Text] [PDF]
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T. Jinquan, C. Jing, H. H. Jacobi, C. M. Reimert, A. Millner, S. Quan, J. B. Hansen, S. Dissing, H.-J. Malling, P. S. Skov, et al. CXCR3 Expression and Activation of Eosinophils: Role of IFN-{gamma}-Inducible Protein-10 and Monokine Induced by IFN-{gamma} J. Immunol., August 1, 2000; 165(3): 1548 - 1556. [Abstract] [Full Text] [PDF]
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C. G. Lerner, M. R. Horton, R. H. Schwartz, and J. D. Powell Distinct Requirements for C-C Chemokine and IL-2 Production by Naive, Previously Activated, and Anergic T Cells J. Immunol., April 15, 2000; 164(8): 3996 - 4002. [Abstract] [Full Text] [PDF]
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K. A. Fitzgerald, A. G. Bowie, B. S. Skeffington, and L. A. J. O'Neill Ras, Protein Kinase C{zeta}, and I{kappa}B Kinases 1 and 2 Are Downstream Effectors of CD44 During the Activation of NF-{kappa}B by Hyaluronic Acid Fragments in T-24 Carcinoma Cells J. Immunol., February 15, 2000; 164(4): 2053 - 2063. [Abstract] [Full Text] [PDF]
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N. E. S. Sibinga, H. Wang, M. A. Perrella, W. O. Endege, C. Patterson, M. Yoshizumi, E. Haber, and M.-E. Lee Interferon-{gamma}-mediated Inhibition of Cyclin A Gene Transcription Is Independent of Individual cis-Acting Elements in the Cyclin A Promoter J. Biol. Chem., April 23, 1999; 274(17): 12139 - 12146. [Abstract] [Full Text] [PDF]
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M. R. Horton, S. Shapiro, C. Bao, C. J. Lowenstein, and P. W. Noble Induction and Regulation of Macrophage Metalloelastase by Hyaluronan Fragments in Mouse Macrophages J. Immunol., April 1, 1999; 162(7): 4171 - 4176. [Abstract] [Full Text] [PDF]
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M. R. Horton, C. M. McKee, C. Bao, F. Liao, J. M. Farber, J. Hodge-DuFour, E. Pure, B. L. Oliver, T. M. Wright, and P. W. Noble Hyaluronan Fragments Synergize with Interferon-gamma to Induce the C-X-C Chemokines Mig and Interferon-inducible Protein-10 in Mouse Macrophages J. Biol. Chem., December 25, 1998; 273(52): 35088 - 35094. [Abstract] [Full Text] [PDF]
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 J. Hodge-Dufour, M. W. Marino, M. R. Horton, A. Jungbluth, M. D. Burdick, R. M. Strieter, P. W. Noble, C. A. Hunter, and E. Pure Inhibition of interferon gamma induced interleukin 12 production: A potential mechanism for the anti-inflammatory activities of tumor necrosis factor PNAS, November 10, 1998; 95(23): 13806 - 13811. [Abstract] [Full Text] [PDF]
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