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*
Department of Microbiology and Immunology, University of Adelaide, Adelaide, Australia; and
Biomedical Research Center, University of British Columbia, Vancouver, British Columbia, Canada
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Abstract |
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 Top Abstract IntroductionMaterials and MethodsResultsDiscussionReferences |
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(8–73), inhibited calcium
mobilization induced by MIP-2, but not by platelet-activating factor in
leukocytes isolated from the bone marrow, indicating that this
antagonist inhibits MIP-2 activity toward murine leukocytes.
Pretreatment of mice with GRO(8–73) inhibited, in a dose-dependent
manner, the MIP-2-induced influx of neutrophils to levels that were not
significantly different from control values. Moreover, this antagonist
was also effective in inhibiting the leukocyte recruitment induced by
TNF-, LPS, and IL-1ß. Leukocyte infiltration into the peritoneal
cavity in response to MIP-2 was also inhibited by prior treatment of
mice with GRO(8–73) or the analogue of platelet factor 4,
PF4(9–70). The results of this study indicate 1) that the murine
receptor for MIP-2 and KC, muCXCR2, plays a major role in neutrophil
recruitment to s.c. tissue and the peritoneal cavity in response to
proinflammatory agents and 2) that CXCR2 receptor antagonists prevent
acute inflammation in vivo. |
Introduction |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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Members of the chemokine gene superfamily of cytokines share homologous sequences and a highly conserved cysteine motif. Chemokines can be divided into four branches, depending on whether the first two cysteines are separated (CXC) or not (CC) by an intervening amino acid (7), whether the second cysteine is missing (C) (8), or whether the first pair of cysteines are separated by three intervening amino acids (CX3C) (9, 10). Members of the CXC subfamily can be divided into two classes: Glu-Leu-Arg-containing CXC chemokines and non-Glu-Leu-Arg-containing CXC chemokines (Glu-Leu-Arg being the three amino acids before the CXC motif). The former are chemotactic in vitro for neutrophils but not for mononuclear cells, whereas the latter exhibit chemotactic activity toward monocytes and lymphocytes (11).
Collectively, chemokines mediate the recruitment of all known
subpopulations of leukocytes. Antagonism of chemokine receptors
therefore represents an important therapeutic target for the treatment
of a variety of disease processes. N-terminal modification of several
chemokines has been shown to antagonize the binding of ligands to the
receptor for such chemokines. For instance, N-terminal deletion
converts monocyte chemoattractant protein-1
(MCP-1)3 and RANTES to
antagonists of native MCP-1 and RANTES, MCP-3 and
macrophage-inflammatory protein (MIP)-1, respectively (12, 13), whereas retention of the initiating methionine converts
RANTES into a receptor antagonist (14). These antagonists
have been shown to prevent entry of HIV into target cells (15, 16) and to prevent development of arthritis in animal models
(17, 18). The results of these previous studies have
provided proof of concept for the use of chemokine receptor antagonists
to block chemokine function in vivo.
Structural studies on CXC chemokines have revealed that the receptor
binding and activation domains are also situated at the N terminus of
the molecules (19, 20). This has led to the development of
truncated analogues of CXC chemokines with the ability to antagonize
CXC receptors. Two such antagonists, growth-related oncogene
(GRO)-8–73(8–73), which comprises aa 8–73, and platelet factor
(PF)-49–70(9–70), which comprises aa 9–70 with an arginine substitution
at the first amino acid, were recently shown to inhibit the binding of
IL-8 and GRO to human CXCR2 receptors as well as the cellular
activation induced by these two chemokines (21). Studies
investigating the role of CXC chemokine receptors in vivo are rare and
to date are limited to examining the effects of genetic deletion
(22). Because in vivo studies are essential to our
understanding of the biologic relevance of various members of the
chemokine gene superfamily, we have investigated the ability of CXC
receptor antagonists to modulate neutrophil recruitment to s.c. tissues
in the mouse in response to the murine CXC chemokine MIP-2, as well as
by other proinflammatory agents. We now report that the truncated
GRO and PF4 analogues function effectively as receptor antagonists
in vivo and that the receptor antagonized, muCXCR2, plays a major role
in neutrophil recruitment to s.c. tissue and the peritoneal
cavity in response to proinflammatory agents.
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Materials and Methods |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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Male BALB/c mice 6–8 wk old were obtained from the Central
Animal House at the University of Adelaide, South Australia. Air
pouches were raised on the dorsum by s.c. injection of 2.5 ml of
sterile air on days 0 and 3. All experiments were conducted on day 6 as
previously described (23). All other reagents were
obtained from the Aldrich Chemical Company (Castle Hill, New South
Wales, Australia). Chemokines and chemokine antagonists were
synthesized as previously described (20). Recombinant
murine TNF- and IL-1ß were purchased from R&D Systems (Jomar
Diagnostics, Magill, South Australia). LPS was purified from
Salmonella enteritidis strain 11RX as previously described
(24). The IgG-purified anti-MIP-2 Abs used in this
study were raised in rabbits using full-length murine (mu) MIP-2 that
was chemically synthesized (23). The Abs were tested for
cross-reactivity against other chemokines (JE, muMIP-1, muMIP-1ß,
muRANTES, C10, TCA-3, and lymphotactin) in direct ELISA and Western
blot. No cross-reactivity was observed.
Induction of leukocyte migration
On day 6, 1 ml of the agonists, as indicated in the figure
legends, dissolved in endotoxin-free PBS or the equivalent volume of
endotoxin-free PBS, was injected in the air pouches. At the indicated
times, the mice were euthanized by asphyxiation with
CO2, the air pouches were washed once with 1 ml
of PBS and then twice with 2 ml of PBS, and the pooled exudates were
centrifuged at 100 x g for 10 min at room temperature.
The supernatants were removed, and the cells were resuspended in PBS,
stained in Turk’s solution (crystal violet 0.01% w/v in acetic acid
3% v/v), and counted. Two hundred thousand cells were centrifuged onto
microscope slides at 500 rpm for 5 min using a cytospin centrifuge
(Shandon, Lab Supply, Adelaide, South Australia). The slides were air
dried and then stained with Diff-Quik (Sigma/Aldrich, Castle Hill, New
South Wales, Australia) to allow quantitation of the granulocyte and
mononuclear leukocyte subpopulations. In air pouch experiments
involving use of GRO8–73(8–73), the indicated concentrations of the
antagonist were injected into the peritoneal cavities the evening
before injection of the agonists.
Induction of peritonitis
One milliliter of either PBS, or GRO8–73(8–73), or PF49–70(9–70)
(both at 250 µg/ml) was injected into the peritoneal cavity of mice.
Five minutes later, 1 ml of either PBS or MIP-2 was injected into the
peritoneal cavity. The mice were sacrificed, and peritoneal cells were
collected and counted 2 h later. Differential analysis was
conducted as described above.
Passive immunization with anti-chemokine Abs
Passive immunization was achieved by injecting 200 µg of either protein G-Sepharose-purified rabbit anti-MIP-2 Abs or the equivalent amount of Ig purified from a preimmune rabbit into the peritoneal cavity of mice the evening before injection of the agonists into the air pouch, as previously described (23).
Measurements of intracellular calcium mobilization
Leukocytes were harvested from the bone marrow of BALB/c mice following dissection of the femur. These cells comprised >85% neutrophils. The cells were suspended at 10 x 106 per ml in HBSS and incubated with 2 µM fura-2-acetoxymethyl ester (fura-2-AM; Molecular Probes, Eugene, OR) for 30 min at 37°C. The cells were washed to remove unincorporated fura-2-AM, and transferred to the thermostated (37°C) cuvette compartment of a fluorometer (Aminco-Bowman AB2, SLM Aminco, Rochester, NY), and the fluorescence was monitored using excitation and emission wavelengths of 340 and 510 nm, respectively. The internal calcium concentrations were calculated as described (25).
Statistical analysis
Numerical values in the figures and tables are means ± SEM. The data for each group were analyzed as indicated in the respective figure legends either by unpaired, two-way ANOVA with a Bonferroni posttest, or using the Student’s t test (unpaired, two-way). Statistical significance was considered to be achieved at p < 0.05.
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Results |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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8–73(8–73) on activation of murine
leukocytes by chemokines
The ability of the CXCR2 antagonist GRO8–73(8–73) to inhibit
murine leukocyte activation by MIP-2 was examined in vitro using the
intracellular calcium mobilization assay. MIP-2 induced a rapid,
transient mobilization of intracellular calcium in cells isolated from
the bone marrow of BALB/c mice, and prior exposure of these cells to
MIP-2 led to homologous desensitization to further treatment with
MIP-2, a characteristic typical of the interaction of chemokines with
their receptors (Fig. 1
A).
GRO8–73(8–73) at a concentration 100-fold that of MIP-2 failed to
induce an increase in the level of intracellular calcium (Fig. 1B). However, prior exposure of the cells to GRO8–73(8–73)
completely inhibited the ability of MIP-2 to induce an increase in the
level of intracellular calcium. Platelet-activating factor (PAF) was
included as an agonist at the end of this experiment as a positive
control to indicate that the cells were still responsive to chemotactic
factors (Fig. 1B). The effect of PAF without prior exposure
of the cells to GRO8–73(8–73) is shown in Fig. 1C. Data
collected over a series of experiments are summarized in Table I. In a series of experiments conducted
on leukocytes isolated from the spleen of IL-5 transgenic mice, a rich
source of eosinophils, GRO8–73(8–73) inhibited intracellular calcium
mobilization in response to MIP-2 without modifying the response to
eotaxin (data not shown).
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A single s.c. air pouch was formed on the backs of BALB/c
mice by injection of sterile air. The effect of MIP-2 on the
recruitment of leukocytes into the pouch was then determined. MIP-2 at
various concentrations was injected into the pouches, and after 2
h, the cellular exudate was collected and counted (Fig. 2A). MIP-2 induced a
dose-dependent increase in the number of cells accumulating in the air
pouch.
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B). Low numbers of leukocytes
were present in the air pouch at time 0. No significant increase in
leukocyte accumulation in the air pouches was detected following
injection of diluent (PBS) alone. In contrast, injection of MIP-2
resulted in a transient influx of leukocytes, with maximal accumulation
at between 2 and 4 h after injection. The levels of the cells in
the pouch exudate had returned almost to baseline by 6 h
postinjection.
The ability of the functionally similar murine CXC chemokine, KC, and
its human homologue GRO, to induce leukocyte recruitment into the
pouch exudate was also examined (Fig. 2C). The effect of
GRO was consistently similar to that of MIP-2, in terms of the
number of leukocytes recruited at the same dose, whereas KC was not as
potent. Although KC induced a statistically significant increase in the
number of leukocytes accumulating in the air pouch exudate (
2-fold
greater than control levels), this was only observed at 10 µg KC, and
a dose of 1 µg KC failed to induce a detectable increase in the
number of leukocytes recruited. Over six independent experiments, MIP-2
and GRO, at concentrations of 0.2 and 1.0 µg/ml, consistently
induced a 5-fold increase in the number of leukocytes accumulating
in the air pouch. Because of the lack of potency observed with KC, and
the fact that GRO is a human chemokine, MIP-2 was used in subsequent
experiments.
To characterize the cellular infiltrate in the air pouches induced by
MIP-2, exudate cells collected from air pouches at 2 h
postinjection with different concentrations of MIP-2 were stained and
counted. The results are expressed as the number of neutrophils,
eosinophils, and monocytes per 100 cells counted (Fig. 3A) and the total number of
neutrophils, eosinophils, and monocytes in the exudate (Fig. 3B). The majority of the cells in the pouch at 2 h
postinjection with PBS were neutrophils, with smaller numbers of
eosinophils and monocytes. On stimulation with MIP-2, the relative
levels of these cells altered. In terms of percentage of total cells,
the number of neutrophils increased, whereas the percentage of
monocytes and eosinophils decreased (Fig. 3A). However, the
overall number of all three cell types increased significantly in
response to MIP-2 (data not shown). Representative photomicrographs of
the cell infiltrate obtained are shown in Fig. 4, A and B,
indicating the increase in the relative number of neutrophils
accumulating in the air pouch on injection of MIP-2. T and B
lymphocytes, as identified by morphology, were not observed at any
stage, regardless of the dose of MIP-2 or the time point assessed. This
was also confirmed by flow cytometric analysis (data not shown).
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8–73(8–73)
Mice were given a single i.p. dose of GRO8–73(8–73) 16 h
before injection of MIP-2 into air pouches. The pouch exudate was
collected 2 h after injection of MIP-2, and the number of cells
was counted. The results of these experiments indicate that prior
treatment of mice with the GRO8–73(8–73) inhibits leukocyte recruitment
into the air pouch in a dose-dependent manner (Fig. 5). There was no significant inhibitory
effect of GRO8–73(8–73) at a dose of 50 µg/mouse; however,
significant inhibition was observed when mice were pretreated with 125
and 250 µg of the antagonist.
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, IL-1ß, or LPS on leukocyte recruitment
into the s.c. air pouch. TNF-, IL-1ß, and LPS were therefore
injected into the air pouch, the exudate was collected 2 h later,
and the number of leukocytes present was determined as described above.
All three inflammatory agonists stimulated the influx of leukocytes
into the air pouch (Table II). The cells
were also differentially stained and enumerated. The results are
expressed as the number of neutrophils, eosinophils, and monocytes per
100 cells counted (Fig. 6A)
and the total number of neutrophils, eosinophils, and monocytes in the
exudate (Fig. 6B). As observed with MIP-2, the major cell
type entering the air pouch on stimulation was the neutrophil, although
this varied between agonists: LPS exclusively induced neutrophil
recruitment, whereas TNF- also increased the recruitment of
monocytes. In contrast, compared with the control values, IL-1ß
induced recruitment of neutrophils and monocytes to approximately the
same extent.
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8–73(8–73) inhibits leukocyte recruitment
induced by TNF-, LPS, and IL-1ß, mice were pretreated with 250
µg of GRO8–73(8–73), and the following day, TNF-, LPS, IL-1ß,
MIP-2, or GRO (the latter as positive controls) was injected into
the air pouches. The exudates were collected 2 h later, and the
number of cells accumulating was determined. Prior treatment of the
mice with GRO8–73(8–73) significantly inhibited leukocyte recruitment
in response to all of the agonists tested (Fig. 7A). As a further control, the
effect of a synthetic CC chemokine receptor antagonist on acute
inflammation induced by TNF- was examined. Prior treatment of mice
with RANTES9–68(9–68) at a dose shown to inhibit development of arthritis
in the MRL/lpr mouse (I. Clark-Lewis, unpublished
observation) failed to inhibit leukocyte recruitment into air pouches
in response to TNF- (Fig. 7B).
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The results obtained using GRO8–73(8–73) suggest that
activation of chemokine receptors including the receptor for MIP-2 and
KC is responsible for the infiltration observed following injection of
TNF-, IL-1ß, and LPS. Thus, it is likely that these are inducing
the production of MIP-2 or related chemokines that also bind to the
same receptors as the GRO8–73(8–73). Abs to MIP-2 were therefore used
to further evaluate the role of MIP-2 in the neutrophil recruitment in
response to TNF-, IL-1, and LPS. Mice were pretreated with
polyclonal Abs directed against MIP-2 or Abs isolated from a preimmune
rabbit 16 h before induction of inflammation (Fig. 8). Mice were also injected with MIP-2 as
a positive control. The exudate cells were collected 2 h
postinjection and counted. The response to all four agonists was
significantly inhibited by the anti-MIP-2 Abs, whereas the
preimmune serum had no detectable effect on the induction leukocyte
recruitment. The inhibition of leukocyte recruitment was observed
equally at the level of neutrophil, eosinophil, and mononuclear
phagocyte recruitment (data not shown).
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8–73(8–73) and
PF49–70(9–70)
In select experiments, the effect of the CXCR2 antagonist on
leukocyte recruitment to the peritoneal cavity was also assessed. In
addition, the ability of a related truncated chemokine antagonist,
PF49–70(9–70), was examined. Mice were treated i.p. with 250 µg of
GRO8–73(8–73) or PF49–70(9–70) 5 min before injection of either PBS or
MIP-2 into the peritoneal cavity. The peritoneal cavities were then
washed as described in Materials and Methods, and
the number of leukocytes accumulating was assessed. Treatment with
either of the antagonists effectively inhibited neutrophil recruitment
in response to MIP-2 (Fig. 9).
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Discussion |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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, muIL-1ß, and LPS.
These results demonstrate that chemokine receptor antagonists block
neutrophil recruitment in vivo and suggest the potential usefulness of
CXCR2 antagonists in control of acute inflammation.
The two antagonists used in the present study, PF49–70(9–70) and
GRO8–73(8–73), have previously been characterized with human
neutrophils (21). Both molecules inhibited functions
mediated by CXCR2, which is activated by IL-8, GRO, and all of the
other Glu-Leu-Arg-containing CXC chemokines (11, 19). In
the human system, IL-8 binds a second private receptor, CXCR1, and the
two receptors, CXCR1 and CXCR2, appear to be coexpressed on neutrophils
and elicit similar functions, although functional differences have been
reported (26, 27, 28). The GRO8–73(8–73) antagonist is a
truncation analogue of human GRO, whereas the PF49–70(9–70) antagonist
is a truncation analogue of human PF4 that contains an N-terminal
arginine substitution (21). The mouse system differs
significantly from the human, as no murine homologue of IL-8 or of
CXCR1 has been identified (22). It is therefore possible
that the functions attributed to IL-8 and CXCR1 in humans are
accomplished in the mouse by other Glu-Leu-Arg-containing CXC
chemokines acting exclusively on CXCR2. Like humans, other primates,
sheep, pigs, and rabbits also have IL-8. Thus, the mouse system
provides a useful model for the in vivo study of antagonists for CXCR2.
The previous results demonstrating receptor antagonism by truncation
mutants suggest that such antagonists may prove useful in determining
the importance of CXCR2 in acute inflammatory responses (12, 13).
A mouse generated by gene targeting that lacks muCXCR2
(22) had defective neutrophil recruitment to the
peritoneal cavity in response to thioglycolate. However, this mouse is
characterized by pronounced neutrophilia, an abnormal production of
myeloid stem cells, and B lymphocytosis. These primary defects
therefore make it difficult to interpret the precise role of CXCR2 in
inflammatory responses. Moreover, chemokines normally act in
inflammatory conditions that are localized to specific tissues and that
usually follow a progressive disease pattern resulting in either
chronic disease or eventual resolution. Thus, whereas the
CXCR2-/- mouse demonstrates the effect of
chronic deficiency in responses mediated by this receptor, it does not
indicate the effects of blocking responses under acute inflammatory
conditions. The present study takes a different approach of inhibiting
CXCR2 function by competition with endogenous or exogenously
administered ligands. CXC chemokines have also previously been
implicated in acute inflammatory responses in vivo in the mouse using
neutralizing Abs (23). However, whereas the Ab approach
targets one ligand, the antagonists used in the present study
target the receptor, which effectively blocks activity of all ligands
on that receptor (12, 13, 21). The results of the present
study suggest that direct antagonism of the receptor utilized by MIP-2
in the murine system, muCXCR2, effectively inhibits neutrophil
recruitment induced by MIP-2 as well as by TNF-, IL-1ß, and LPS,
and therefore provide the clearest evidence to date of the importance
of CXCR2 in neutrophil recruitment in the murine system under acute
inflammatory conditions.
Previous data have demonstrated the relative importance of both KC and
MIP-2, two CXCR2 ligands, in the mouse in terms of neutrophil
recruitment into s.c. air pouches in response to TNF-
(23). Passive immunization of mice with neutralizing Abs
directed against either KC or MIP-2 effectively inhibited neutrophil
accumulation in response to TNF- as well as by IL-1ß and LPS (the
present study), supporting the notion that neutrophil recruitment in
response to the latter inflammatory agents is due in large part to the
production of these molecules. In the present study, it was observed
that MIP-2 was far more potent than KC in inducing leukocyte
accumulation in the air pouch. Although recombinant KC has been shown
to be active as a chemoattractant for murine neutrophils in vitro, a
similar low potency was reported for neutrophil accumulation in the
peritoneal cavity in response to KC (29). Assuming that
the muKC used in both of these studies was fully active, these combined
data suggest that KC may not play as important a role in active
neutrophil recruitment to s.c. tissue as MIP-2. Indeed, in that
previous study, KC was reported to be constitutively expressed in s.c.
tissue, leading to the hypothesis that it may play an important role in
basal trafficking of neutrophils (29).
In contrast to the human situation, in which neutrophil migration is
induced by ligands for CXCR1 or CXCR2, in the mouse, the CC chemokine
MIP-1 also induces neutrophil migration through interaction with its
receptor muCCR1 (23, 30, 31, 32). We found in the present
study that the CXCR2 antagonist did not inhibit the ability of
muMIP-1 to induce activation of murine leukocytes as determined in
the calcium mobilization assay (data not shown), indicating that this
antagonist does not interfere with the binding of muMIP-1 to its
receptor(s). The fact that all of the neutrophil responses could be
substantially (indeed, almost completely) blocked by CXCR2 antagonists,
despite the known involvement of MIP-1 in neutrophil recruitment,
may be explained in several ways: first, that muCXCR2 ligands are
dominant in the models of acute inflammatory responses used in the
present study; second, that muCXCR2 and muCCR1 are both required for
neutrophil recruitment in these models, and blockade of one of these
receptors effectively inhibits all recruitment; and third, that because
MIP-1 is produced upon mitogenic stimulation of T cells (6, 33), it could be more important as a neutrophil chemoattractant
in inflammatory responses involving T cells, rather than in the acute
inflammatory responses investigated in the present study. In a previous
study, we showed that inhibition by neutralizing Abs of the activity of
endogenously synthesized MIP-1 inhibits neutrophil recruitment into
s.c. air pouches during acute inflammation induced by exogenous TNF-
(23). Thus, the second theory appears to be correct.
Although these previous data point to a role for MIP-1 in neutrophil
recruitment into the s.c. air pouch, it is important to note that
direct injection of this CC chemokine does not induce accumulation of
neutrophils (23). These combined observations imply the
existence of a chemokine network in acute inflammation, involving
several chemokines of which the roles appear to be regulated
temporally. Inhibiting the biologic activity of one of several
chemokines in the network therefore inhibits the biologic response.
Such a theory appears to explain the consistent ability of Abs directed
against single chemokines to significantly inhibit biologic effects in
vivo when other chemokines are known or suspected to play a
similar role.
In summary, our data indicate that CXCR2 antagonists, which had been
previously characterized in vitro, also function effectively in vivo.
The results of this study provide several important pieces of
information. First, they indicate that the receptor antagonized by
GRO8–73(8–73) and PF49–70(9–70), muCXCR2, is a major receptor mediating
neutrophil recruitment to s.c. tissue in response to direct injection
of MIP-2, as well as following injection of IL-1ß, TNF-, and LPS,
agonists that act via the induction of MIP-2. Second, while it is clear
that, because of a lack of potency, the CXC truncation mutants studied
in this report will not be useful therapeutically, our data provide
proof of concept that small molecule CXCR antagonists may be useful in
controlling acute inflammation by blocking neutrophil recruitment.
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Acknowledgments |
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Footnotes |
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2 Address correspondence and reprint requests to Dr. Shaun R. McColl, Chemokine Biology, Department of Microbiology and Immunology, University of Adelaide, Adelaide 5005, Australia. E-mail address:
3 Abbreviations used in this paper: MCP-1, monocyte chemoattractant protein-1; MIP, macrophage-inflammatory protein; GRO, growth-related oncogene; PF, platelet factor; mu, murine; fura-2-AM, fura-2-acetoxymethyl ester; PAF, platelet-activating factor.
Received for publication June 16, 1998. Accepted for publication June 25, 1999.
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 T. H. Thatcher, N. A. McHugh, R. W. Egan, R. W. Chapman, J. A. Hey, C. K. Turner, M. R. Redonnet, K. E. Seweryniak, P. J. Sime, and R. P. Phipps Role of CXCR2 in cigarette smoke-induced lung inflammation Am J Physiol Lung Cell Mol Physiol, August 1, 2005; 289(2): L322 - L328. [Abstract] [Full Text] [PDF]
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K. Arora, W. M. Gwinn, M. A. Bower, A. Watson, I. Okwumabua, H. R. MacDonald, M. I. Bukrinsky, and S. L. Constant Extracellular Cyclophilins Contribute to the Regulation of Inflammatory Responses J. Immunol., July 1, 2005; 175(1): 517 - 522. [Abstract] [Full Text] [PDF]
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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]
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J. A. Lekstrom-Himes, D. B. Kuhns, W. G. Alvord, and J. I. Gallin Inhibition of Human Neutrophil IL-8 Production by Hydrogen Peroxide and Dysregulation in Chronic Granulomatous Disease J. Immunol., January 1, 2005; 174(1): 411 - 417. [Abstract] [Full Text] [PDF]
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C. Hess, T. K. Means, P. Autissier, T. Woodberry, M. Altfeld, M. M. Addo, N. Frahm, C. Brander, B. D. Walker, and A. D. Luster IL-8 responsiveness defines a subset of CD8 T cells poised to kill Blood, December 1, 2004; 104(12): 3463 - 3471. [Abstract] [Full Text] [PDF]
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 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]
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K. R. Pilkington, I. Clark-Lewis, and S. R. McColl Inhibition of Generation of Cytotoxic T Lymphocyte Activity by a CCL19/Macrophage Inflammatory Protein (MIP)-3{beta} Antagonist J. Biol. Chem., September 24, 2004; 279(39): 40276 - 40282. [Abstract] [Full Text] [PDF]
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J. Godau, T. Heller, H. Hawlisch, M. Trappe, E. Howells, J. Best, J. Zwirner, J. S. Verbeek, P. M. Hogarth, C. Gerard, et al. C5a Initiates the Inflammatory Cascade in Immune Complex Peritonitis J. Immunol., September 1, 2004; 173(5): 3437 - 3445. [Abstract] [Full Text] [PDF]
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 S. T. Tarzami, W. Miao, K. Mani, L. Lopez, S. M. Factor, J. W. Berman, and R. N. Kitsis Opposing Effects Mediated by the Chemokine Receptor CXCR2 on Myocardial Ischemia-Reperfusion Injury: Recruitment of Potentially Damaging Neutrophils and Direct Myocardial Protection Circulation, November 11, 2003; 108(19): 2387 - 2392. [Abstract] [Full Text] [PDF]
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 L.-Y. Chen, J.-J. Shieh, B. Lin, C.-J. Pan, J.-L. Gao, P. M. Murphy, T. F. Roe, S. Moses, J. M. Ward, E. J. Lee, et al. Impaired glucose homeostasis, neutrophil trafficking and function in mice lacking the glucose-6-phosphate transporter Hum. Mol. Genet., October 1, 2003; 12(19): 2547 - 2558. [Abstract] [Full Text] [PDF]
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A. L. Miller, R. M. Strieter, A. D. Gruber, S. B. Ho, and N. W. Lukacs CXCR2 Regulates Respiratory Syncytial Virus-Induced Airway Hyperreactivity and Mucus Overproduction J. Immunol., March 15, 2003; 170(6): 3348 - 3356. [Abstract] [Full Text] [PDF]
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P. L. Podolin, B. J. Bolognese, J. J. Foley, D. B. Schmidt, P. T. Buckley, K. L. Widdowson, Q. Jin, J. R. White, J. M. Lee, R. B. Goodman, et al. A Potent and Selective Nonpeptide Antagonist of CXCR2 Inhibits Acute and Chronic Models of Arthritis in the Rabbit J. Immunol., December 1, 2002; 169(11): 6435 - 6444. [Abstract] [Full Text] [PDF]
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E.-B. Haddad, K. McCluskie, M. A. Birrell, D. Dabrowski, M. Pecoraro, S. Underwood, B. Chen, G. T. De Sanctis, S. E. Webber, M. L. Foster, et al. Differential Effects of Ebselen on Neutrophil Recruitment, Chemokine, and Inflammatory Mediator Expression in a Rat Model of Lipopolysaccharide-Induced Pulmonary Inflammation J. Immunol., July 15, 2002; 169(2): 974 - 982. [Abstract] [Full Text] [PDF]
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 W. I. de Boer Cytokines and Therapy in COPD* : A Promising Combination? Chest, May 1, 2002; 121(5_suppl): 209S - 218S. [Abstract] [Full Text] [PDF]
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M. T. Wiekowski, S.-C. Chen, P. Zalamea, B. P. Wilburn, D. J. Kinsley, W. W. Sharif, K. K. Jensen, J. A. Hedrick, D. Manfra, and S. A. Lira Disruption of Neutrophil Migration in a Conditional Transgenic Model: Evidence for CXCR2 Desensitization In Vivo J. Immunol., December 15, 2001; 167(12): 7102 - 7110. [Abstract] [Full Text] [PDF]
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A. M. Hohlbaum, M. S. Gregory, S.-T. Ju, and A. Marshak-Rothstein Fas Ligand Engagement of Resident Peritoneal Macrophages In Vivo Induces Apoptosis and the Production of Neutrophil Chemotactic Factors J. Immunol., December 1, 2001; 167(11): 6217 - 6224. [Abstract] [Full Text] [PDF]
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S. P. Matzer, T. Baumann, N. W. Lukacs, M. Rollinghoff, and H. U. Beuscher Constitutive Expression of Macrophage-Inflammatory Protein 2 (MIP-2) mRNA in Bone Marrow Gives Rise to Peripheral Neutrophils with Preformed MIP-2 Protein J. Immunol., October 15, 2001; 167(8): 4635 - 4643. [Abstract] [Full Text] [PDF]
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U. Baumann, N. Chouchakova, B. Gewecke, J. Kohl, M. C. Carroll, R. E. Schmidt, and J. E. Gessner Distinct Tissue Site-Specific Requirements of Mast Cells and Complement Components C3/C5a Receptor in IgG Immune Complex-Induced Injury of Skin and Lung J. Immunol., July 15, 2001; 167(2): 1022 - 1027. [Abstract] [Full Text] [PDF]
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N. J. Coates and S. R. McColl Production of Chemokines In Vivo in Response to Microbial Stimulation J. Immunol., April 15, 2001; 166(8): 5176 - 5182. [Abstract] [Full Text] [PDF]
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N. Chouchakova, J. Skokowa, U. Baumann, T. Tschernig, K. M. H. Philippens, B. Nieswandt, R. E. Schmidt, and J. E. Gessner Fc{{gamma}}RIII-Mediated Production of TNF-{{alpha}} Induces Immune Complex Alveolitis Independently of CXC Chemokine Generation J. Immunol., April 15, 2001; 166(8): 5193 - 5200. [Abstract] [Full Text] [PDF]
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 K. Tateda, T. A. Moore, M. W. Newstead, W. C. Tsai, X. Zeng, J. C. Deng, G. Chen, R. Reddy, K. Yamaguchi, and T. J. Standiford Chemokine-Dependent Neutrophil Recruitment in a Murine Model of Legionella Pneumonia: Potential Role of Neutrophils as Immunoregulatory Cells Infect. Immun., April 1, 2001; 69(4): 2017 - 2024. [Abstract] [Full Text] [PDF]
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E. J. Kunkel, J. L. Dunne, and K. Ley Leukocyte Arrest During Cytokine-Dependent Inflammation In Vivo J. Immunol., March 15, 2000; 164(6): 3301 - 3308. [Abstract] [Full Text] [PDF]
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H. Shinohara, S. Yano, C. D. Bucana, and I. J. Fidler Induction of Chemokine Secretion and Enhancement of Contact-Dependent Macrophage Cytotoxicity by Engineered Expression of Granulocyte-Macrophage Colony-Stimulating Factor in Human Colon Cancer Cells J. Immunol., March 1, 2000; 164(5): 2728 - 2737. [Abstract] [Full Text] [PDF]
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