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(Received for publication, June 28, 1995) From the
Interleukin 8 (IL-8) is a potent chemoattractant and activating
factor for human polymorphonuclear leukocytes (PMN) and hence plays a
critical role in the pathogenesis of acute inflammation. Two unique but
homologous receptors for IL-8 have been cloned (IL-8RA and -B), each of
which binds the IL-8 ligand with high affinity. PMN stimulated by
cytokines or lipopolysaccharide (LPS) exhibit changes in IL-8R mRNA and
The recruitment and activation of polymorphonuclear leukocytes
is the hallmark of acute inflammation. Chemotactic factors produced at
an inflammatory site regulate vascular adhesion, transendothelial
migration, and the movement of leukocytes through the extracellular
matrix. Interleukin 8 is the prototype of a family of chemoattractant
cytokines known as chemokines(1) , which regulate these
migratory processes and determine the cellular composition of the
inflammatory response by their target cell specificity (reviewed in
Refs. 1 to 3). In particular, several members of the chemokine Two receptors exhibiting high affinity
binding (K In
order to better understand the physiological control of IL-8-mediated
events in acute inflammation we have investigated the in vitro regulation of the IL-8 receptors A and B in human PMN.
Figure 1:
Regulation of IL-8R mRNA expression in
PMN. RNA samples (5 µg per lane) were prepared from purified
peripheral blood PMN incubated for 6 h in the presence of various
stimuli. Lanes from left to right, medium alone (RPMI/10%
fetal calf serum), IL-8 (50 ng/ml), TNF-
Figure 2:
A, kinetics of IL-8R regulation in
peripheral blood PMN induced by G-CSF and LPS. RNA samples (5
µg/lane) were prepared from purified peripheral blood PMN incubated
for 1-18 h in the presence or absence of G-CSF (50 ng/ml) or LPS
(10 ng/ml). The autoradiographic signals were measured by densitometry
and normalized to
Figure 3:
Nuclear run-on analysis of the IL-8R mRNA
in PMN treated with G-CSF or LPS. Nuclei were prepared from 2-5
Figure 4:
Half-life of the IL-8R mRNA in PMN treated
with LPS. RNA samples (5 µg/lane) were prepared from purified
peripheral blood PMN incubated for 1-18 h: with medium alone,
with actinomycin D (10 µM), with LPS (10 ng/ml), or
preincubated with actinomycin D (10 µM) for 30 min
followed by the addition of LPS (10 ng/ml). The autoradiographic
signals were measured by densitometry and normalized to those of
Figure 5:
Regulation of IL-8 binding and
IL-8-induced chemotaxis on peripheral blood PMN by G-CSF or LPS.
Purified peripheral blood PMN were incubated for 1-20 h in the
presence or absence of G-CSF (50 ng/ml) or LPS (10 ng/ml) before being
included in standard
The IL-8RA and -B are abundantly expressed on human PMN. The
IL-8R mRNAs are reciprocally regulated in response to stimuli such as
G-CSF and LPS, which are likely to be present in sites of acute
inflammation. Responses to chemoattractant cytokines, including
IL-8, may be desensitized by continued stimulation(22) ,
confirming that the chemokine receptors are regulated via
agonist-dependent mechanisms as has been shown for other rhodopsin
family members (reviewed in (23) ). In the present work, we
have demonstrated that the two IL-8R also exhibit regulation in
response to exogenous stimuli. Treatment of PMN with G-CSF resulted in
increased IL-8R mRNA expression, as well as IL-8 binding and
chemotactic responsiveness. G-CSF has been shown to enhance PMN
survival in vitro by inhibition of apoptosis(24) .
However, the up-regulation of IL-8R expression we have demonstrated is
not explicable simply on the basis of improved PMN survival, as the
IL-8R mRNA and chemotactic response to IL-8 are significantly increased
above baseline levels in G-CSF-treated PMN ( Fig. 2and Fig. 5). G-CSF enhances IL-8R expression by a transcriptional
mechanism, as actinomycin D treatment blocked its effect, and nuclear
run-on studies showed increased IL-8R signals after G-CSF treatment of
PMN. This regulatory pathway, if direct, may indicate a novel
G-CSF-responsive transcriptional regulatory element in the IL-8R genes.
Indeed, we have recently isolated, sequenced, and characterized the
genomic structure of the IL-8RB gene(25) . IL-8RB promoter
region-CAT constructs demonstrated enhanced expression in HL60 cells
after treatment with G-CSF, thus providing evidence for a direct
transcriptional effect of G-CSF on the IL-8RB gene. Comparison of the
promoter region of this gene with that of another PMN chemoattractant
receptor, the fMLP receptor, has demonstrated the presence of several
novel, but highly conserved, sequence motifs, which may represent
tissue-specific transcriptional regulatory elements(25) .
Interestingly, these motifs were not detected in the promoter region of
the IL-8RA gene(26) . Down-regulation of IL-8R expression
and IL-8 responsiveness was demonstrated when PMN were incubated with
LPS. This treatment induced a significant decrease in the half-life of
the IL-8R mRNA, consistent with an effect on the stability of the
message. In addition, IL-8R transcriptional activity appeared to be
inhibited in nuclear run-on studies. LPS also has been shown to enhance
the survival of PMN in vitro(24) , and to induce the
expression of several genes including IL-1 The inhibitory effect of LPS
on the chemotactic response of PMN to IL-8 was detected earlier than
expected from the alteration in the mRNA. This may have been
attributable in part to clumping and adhesion of PMN to the upper side
of the polycarbonate filter. We have observed similar clumping effects
with fMLP, yet LPS treatment enhances fMLP receptor mRNA expression in
PMN and augments the PMN chemotactic response to fMLP. ( Our findings suggest that stimuli occurring in
vivo in acute inflammation may alter the expression of IL-8R on
PMN. We have provided preliminary evidence for the functional
significance of this regulation in PMN chemotaxis. The capacity of
G-CSF to stimulate PMN functions, including phagocytosis and superoxide
generation, may be mediated in part by enhanced responsiveness to IL-8.
Interestingly, exposure of PMN to LPS may result not only in the
synthesis and release of a secondary cascade of proinflammatory
cytokines including IL-1
Volume 270,
Number 47,
Issue of November 24, 1995 pp. 28188-28192
©1995 by The American Society for Biochemistry and Molecular Biology, Inc.
ABSTRACT
INTRODUCTION
MATERIALS AND METHODS
RESULTS
DISCUSSION
FOOTNOTES
ACKNOWLEDGEMENTS
REFERENCES
I-IL-8 binding. Granulocyte-colony stimulating factor
(G-CSF) treatment of PMN enhances, and LPS inhibits, IL-8R mRNA
expression. Similarly,
I-IL-8 ligand binding to PMN is
increased by G-CSF and decreased by LPS treatment. The stimulatory
effect of G-CSF on IL-8R expression is transcriptional as it is
inhibited by actinomycin D and is evident in nuclear run-on analyses.
In contrast, LPS down-regulates IL-8R by both transcriptional and
post-transcriptional mechanisms. The alterations in IL-8R expression
are associated with similar changes in the IL-8-induced chemotactic
responses of PMN. In conclusion, the two types of IL-8 receptor differ
in their cellular distribution and are regulated in response to
cytokines and LPS. Regulation of IL-8R expression by endogenous and
exogenous immunomodulators may be important in the in vivo control of PMN effector functions in inflammation.
subfamily: IL-8, (
)GRO/melanoma growth-stimulating activity,
neutrophil-activating peptide 2 (NAP-2), and ENA-78 are neutrophil
chemoattractants(4, 5, 6, 7) . These
chemokines are produced by a wide range of cell types, notably
macrophages, in response to a diverse array of stimuli including
proinflammatory cytokines, such as IL-1 and TNF, as well as
LPS(1, 2) .
2 nM) for IL-8
have been cloned, and hence have been designated as type A and B IL-8
receptors. The type A IL-8 receptor was cloned by direct expression
from a PMN library (8) and binds IL-8(9, 10) .
In contrast, the type B receptor was identified from HL60
myelomonocytic cells (11) and has subsequently been
demonstrated to also bind GRO-/melanoma growth-stimulating
activity with high affinity (K
2
nM), and NAP-2 and ENA-78 with lower
affinity(9, 10) . The IL-8R proteins are members of
the rhodopsin superfamily of seven-transmembrane domain,
G-protein-coupled receptors. They share 77% overall amino acid
identity, including two matching regions of 105 and 64 amino acids, and
their genes co-localize to chromosome 2q35 (12) . The chemokine
receptors, including the IL-8R and the recently cloned 
chemokine
receptors(13, 14) , form a new subfamily of the
rhodopsin receptor superfamily. IL-8RA is strongly expressed only in
PMN, although the mRNA is detectable in differentiated HL60 cells (15) . In contrast, IL-8RB is prominently expressed in PMN, and
the mRNA is also widely distributed in myelomonocytic cell lines (U937,
THP-1, and HL60), the Jurkat T cell line, as well as melanoma and
fibroblast lines(15) . In addition, we have recently obtained
evidence that the IL-8R mRNAs are expressed in freshly prepared
peripheral blood T cells, but are progressively diminished by
incubation with or without anti-CD3 stimulation(16) .
Receptor cDNAs
A 1050-base pair sequence of the
IL-8RA cDNA (nucleotides -20-1030 of the published cDNA)
was amplified in a polymerase chain reaction with two synthetic
oligonucleotides, 5`-ttgctgaaactgaagaa-3`, 5`-gacattgacagacgaaga-3`,
from human genomic DNA using a thermocycler oven (Bios Corp., New
Haven, CT). The reaction conditions consisted of three cycles of 96
°C 
1 min, 65 °C 1 min, 72 °C 1 min,
followed by 30 cycles of 94 °C 30 s, 65 °C 45
s, 72 °C 30 s, and a final extension step of 72 °C
10 min. For Northern analyses, cDNAs encoding regions of
divergence between IL-8RA and -B were prepared. A 245-base pair AccI fragment (nucleotides 1-245) of this IL-8RA cDNA
and a 552-base pair HhaI-HindIII fragment of an
IL-8RB clone previously isolated from a U937 myelomonocytic cell
library (Invitrogen) (12) consisting of nucleotides 698-1250 of
the published cDNA were chosen. These regions share only approximately
60% nucleotide homology with the analogous region of the other IL-8R.Cells
Peripheral blood leukocytes enriched for
mononuclear cells or for granulocytes were obtained from normal donors
by leukapheresis. Granulocytes were purified by dextran sedimentation,
followed by Ficoll gradient centrifugation and hypotonic lysis of red
blood cells. PMN were collected, washed in phosphate-buffered saline,
and resuspended at 2-5 10
/ml in RPMI 1640
supplemented with 10% fetal calf serum. The purity of the PMN
preparations was judged to be greater than 90-95% by
morphological criteria; the remaining cells were typically lymphocytes. Cell Stimulation
Purified PMN were incubated in
tissue culture flasks (Costar) in the presence or absence of various
cytokines or LPS. Prior to harvesting for RNA preparation, binding or
chemotaxis assays cell viability was assessed by trypan blue exclusion.
The recombinant human cytokines used in these studies were obtained
from the following sources: IL-8 was provided by Dr. M. Yamada of
Dainippon Pharmaceutical Co. Ltd. (Osaka, Japan). TNF-, GM-CSF,
G-CSF, and GRO- were provided by Dr. Craig Reynolds of the
National Cancer Institute (Frederick, MD). The recombinant G-CSF was
originally obtained from Amgen (Thousand Oaks, CA). Purified LPS (Escherichia coli 055:B5) was purchased from Difco
Laboratories. Actinomycin D and fMLP were purchased from Sigma.
Purified peripheral blood T cells were activated for 8 h on anti-CD3
(OKT3, Becton Dickinson) coated plates prior to harvesting for RNA.Northern Blot Analyses
Total RNA was prepared from
guanidinium thiocyanate-treated cell lysates using CsCl
ultracentrifugation(17) . The RNA was electrophoresed through
1% agarose/formaldehyde gels. The samples were transferred to nylon
membranes (Nytran, Schleicher & Schuell), fixed by UV
cross-linking, and hybridized overnight in Hybrisol I (Oncor,
Gaithersburg, MD) at 42 °C with 1-3 10 cpm/ml P-labeled cDNA probe. Filters were washed at
65 °C in 0.1
SSC/0.1% SDS and then exposed to Kodak XAR5
film for 1-7 days. Bands on the autoradiograms were integrated
with a densitometer (LKB Ultroscan), and values were normalized to the
signal of a control -actin cDNA.Nuclear Run-on Assays
Nuclear run-on analyses were
performed as described previously (18) with the following
modifications. Nuclei were prepared from 2-5 10
PMN per variable by hypotonic and Nonidet P-40 lysis followed by
sucrose gradient centrifugation. Each labeling reaction included 150
µCi of [-P]UTP (800 Ci/mmol, Amersham)
in a buffer containing 139 mM KCl, 10 mM Tris-HCl, pH
8.0, 5 mM dithiothreitol, 0.2 mM EDTA, 0.8 mM each of dATP, dCTP, and dGTP. After two rounds of ethanol
precipitation, the RNA pellets were resuspended in TES buffer (10
mM Tris-HCl, pH 7.4, 10 mM EDTA, and 0.2% SDS) before
estimation of the percentage of incorporation into the RNA (usually
70-80%). 10 µg of denatured plasmid cDNAs were slot-blotted
onto nylon membranes, including full-length cDNAs for the IL-8RA and
-B,
-actin (as above), and pGEM-3 as a negative control.
Hybridization was performed for 48-72 h at 42 °C in a freshly
prepared solution (50% deionized formamide, 4 SSC, 0.1% SDS,
100 µg of sonicated salmon sperm DNA, 5 Denhardt's
solution, 100 mM Na
PO
, pH 7.0)
containing 1-2 10
cpm/ml of labeled RNA. The
membrane strips were washed three times for 20 min at room temperature
in 2 SSC, 0.2% SDS, followed by three washes for 30 min at 56
°C in 0.1 SSC, 0.1% SDS. The membranes were exposed to
Kodak XAR5 film for 1-2 weeks after which the autoradiographs
were scanned by densitometer.Receptor Binding Assays
I-IL-8
binding was performed as described previously(19) . In brief, 2
10 PMN were washed twice in chilled
phosphate-buffered saline, resuspended in chilled RPMI 1640 with 10
mg/ml bovine serum albumin (Sigma), 25 mM HEPES, and then
incubated in duplicate with 0.1 ng of I-IL-8 (specific
activity 2200 Ci/mmol; DuPont NEN) in a total volume of 200 µl. The
degree of nonspecific binding was determined by parallel incubation in
the presence of a 500-fold excess of unlabeled IL-8. After incubation
at 4 °C for 2 h, the cells were pelleted by brief centrifugation
and then washed twice with excess binding medium (as above). The tips
of the tubes were cut, and radioactivity was counted in a
counter. The nonspecifically bound radioactivity was subtracted from
the total bound activity to establish specific binding.
Chemotaxis Assays
Cell migration was evaluated
using a 48-well microchemotaxis chamber technique (20) with
polycarbonate filters (5 µm pore size, Nucleopore Corp.,
Pleasanton, CA). PMN were resuspended in RPMI 1640 containing 1% bovine
serum albumin at 1 10 cells/ml, and 50 µl per
well was applied to the upper chamber. Chemoattractants were
resuspended in the same medium and applied to the lower chamber. All
responses were assayed in triplicate. Migration was allowed to continue
at 37 °C for 30 min in a 5% CO, moist atmosphere. After
the nonmigrating cells were washed from the upper surface of the
filters, migrating cells were fixed in methanol and stained with
Diff-Quik. The number of migrating cells in five high-powered fields
were counted for each well.
The IL-8RA and -B mRNA Transcripts
In Northern
analyses of total RNA prepared from PMN, the specific probes showed
that the mRNA transcript for the IL-8RA was 2.4 kilobases and for
IL-8RB was 3.1 kilobases. In addition, in some experiments, the IL-8RA
probe hybridized at high stringency to an additional band at 2.2
kilobases, which may represent a transcript from the IL-8R pseudogene
(designated IL-8RAP; (21) ).Regulation of IL-8R mRNA Expression by Cytokines and
LPS
In initial experiments, purified human PMN were incubated
for 6 h with a panel of different cytokines known to be present in
acute inflammatory states, as well as the neutrophil chemoattractant
fMLP, or with LPS. After each treatment, the viability of the cells was
confirmed to be greater than 95% by Trypan Blue exclusion before
harvesting for RNA. The Northern blots containing these RNAs were
hybridized sequentially with the IL-8R probes, followed by the
-actin probe to assess the relative amounts of RNA loaded. This
analysis demonstrated a substantial increase in specific mRNA for both
IL-8R after treatment with G-CSF at 50 ng/ml and a dramatic reduction
in the steady state IL-8R mRNA after PMN were treated with LPS at 10
ng/ml. A less marked down-regulation was found after stimulation with
tumor necrosis factor (Fig. 1). The G-CSF and LPS effects
were chosen for further study.
(10 ng/ml), fMLP (100
nM), LPS (10 ng/ml), GM-CSF (50 ng/ml), and G-CSF (50 ng/ml).
The autoradiographic signals were measured by densitometry and
normalized to those of -actin. The figure represents one of three
replicate experiments with the quantitative data (upper panel)
and the Northern analysis (lower
panel).
G-CSF and LPS Regulate IL-8R mRNA via Transcriptional
Mechanisms
Changes in the steady-state level of RNA expression
of a gene result from alterations in either the rate of transcription,
the rate of degradation of the message, or a combination of both. After
establishing the optimal doses of G-CSF and LPS (data not shown), the
kinetics of the alterations in the steady state levels of IL-8R
expression in PMN were investigated further. The mRNA expression of
IL-8RA and -B was regulated in parallel in all experiments. Treatment
of PMN with G-CSF for periods up to 18 h produced a dramatic increase
(2-10-fold in four separate experiments) in steady-state IL-8R
mRNA with a plateau reached at approximately 6-18 h. This
contrasted with a slow but steady decline in the mRNA levels in
untreated PMN (Fig. 2A). LPS treatment induced a
significantly more rapid decline in the steady-state mRNA, evident
after 1-2 h of treatment. Actinomycin D at a final concentration
of 10 µM was added to PMN incubations in order to inhibit
transcription. This treatment abrogated the up-regulation of IL-8R mRNA
induced by G-CSF (Fig. 2B), whereas the addition of
actinomycin D to untreated PMN induced a further decline in steady
state mRNA levels. Nuclear run-on analyses confirmed the impression
that G-CSF regulates IL-8R expression by enhancing transcription. The
nuclear RNA signals for IL-8RA and -B in PMN were enhanced
4-6-fold in three separate experiments after 3-6 h of G-CSF
treatment (Fig. 3). These data suggest that IL-8R mRNA
expression in PMN is dependent on continuous transcription of the IL-8R
genes, and, further, that the G-CSF effect is dependent upon enhanced
transcriptional activity of the IL-8R genes.
-actin. The figure represents quantitative data
from one of three replicate experiments. A Northern blot containing
data representative of the LPS effect is included with Fig. 4. B, effect of actinomycin D on the regulation of IL-8R mRNA
expression in PMN by G-CSF. RNA samples (5 µg/lane) were prepared
from purified peripheral blood PMN incubated for 1-18 h with
G-CSF (50 ng/ml) or preincubated for 30 min with actinomycin D (10
µM) followed by the addition of G-CSF (50 ng/ml). The
effect of actinomycin D on IL-8R mRNA regulation by LPS is shown in Fig. 4.
10 purified peripheral blood PMN immediately after
incubation for 3 or 6 h in medium alone (RPMI 1640, 10% fetal calf
serum), or with G-CSF (50 ng/ml) or LPS (10 ng/ml), or a combination of
both stimuli. The labeled nuclear RNA was hybridized with denatured
plasmid cDNAs of IL-8RA and -B, -actin, and pGEM-1. The
autoradiographic signals were measured by densitometry. For
quantitative analyses, the signals from nonspecific hybridization (with
pGEM-1) were subtracted from those of IL-8RA and -B and then normalized
to the signals of -actin. The figure represents one of three
replicate experiments.
-actin. The upper panel is quantitative data from three
replicate experiments (mean ± S.E.). A set of Northern blots
from a representative experiment are included in the lower
panel.
LPS De-stabilizes IL-8R mRNA
The half-life (t) of the IL-8R mRNA was determined by
treatment of PMN in the presence or absence LPS after prior incubation
with actinomycin D. Least squares regression analysis of the normalized
autoradiographic data was used to calculate the numerical value of the t
. In untreated PMN, the IL-8R mRNA t
was 3.6 h (S.E. 0.6 h; n = 3),
whereas in the presence of LPS the half-life was reduced to
approximately 2.2 h (S.E. 0.4 h, n = 3; Fig. 4).
The estimated half-lives for the IL-8RA and -B mRNAs in these
experiments were not significantly different. The nuclear run-on
analyses also suggested that LPS inhibited IL-8R transcription within 3
h of treatment (2-3-fold in three separate experiments; Fig. 3). Furthermore, the addition of LPS, in combination with
G-CSF treatment of PMN, abrogated the up-regulation associated with
G-CSF, resulting in a similar down-regulation of transcriptional
activity to that seen with LPS treatment alone (Fig. 3). Taken
together, these data suggest that LPS inhibits IL-8R expression by a
combination of transcriptional inhibition and decreasing mRNA
stability.
G-CSF and LPS Also Regulate IL-8 Receptor Binding on
PMN
The expression of IL-8R on the surface of PMN was estimated
by measurement of I-IL-8 binding. As both IL-8R
demonstrate similar high affinity for the IL-8 ligand, these data are
likely to reflect the combined binding capacity of both IL-8R types on
the PMN surface. Fig. 5A shows that G-CSF treatment produced
a sustained increase in IL-8 binding which was approximately 2-fold
greater after 20 h in treated PMN in comparison with untreated cells.
In contrast, LPS treatment rapidly reduced IL-8 binding on PMN.
I-IL-8 binding assays performed at 4
°C in the presence or absence of excess unlabeled ligand (A). Specific counts were determined by subtracting the counts
from binding in the presence of excess unlabeled ligand from the total
bound radioactivity by chemotaxis assays, utilizing a modified Boyden
chamber technique, and various doses of IL-8 or medium alone (B). The data represent the mean number of cells per high
power field (±S.D.) detected in 5 fields for the optimal
chemotactic dose of IL-8 (50 ng/ml).
G-CSF and LPS Regulate IL-8 Chemotactic Responses in
PMN
The ability of PMN to migrate in response to IL-8 in a
standard chemotaxis assay was studied in order to examine the possible
functional consequences of the alterations in IL-8R expression. G-CSF
treatment produced a significant increase in the number of PMN
migrating in response to an IL-8 chemotactic gradient (Fig. 5B). LPS treatment was associated with a rapid
reduction in the IL-8 chemotactic response. Similar results were
obtained when GRO- was used as the chemotactic stimulus (data not
shown), confirming that the expression and functional response of
IL-8RB (which binds GRO) is regulated in this system.
(27) and
TNF-(28) . Hence, the inhibitory effect of LPS on IL-8R
expression contrasts with these findings and is not related to reduced
PMN survival. Hormone-induced reduction in mRNA stability has been
documented for several G-protein-coupled receptors including the
-adrenergic receptor and thyrotropin-releasing hormone
receptor(23) . Although the mechanisms of this altered RNA
stability are not yet clear, the 3`-untranslated region of the mRNA for
the -adrenergic receptor, as well as several other
rhodopsin family members, contain AU-rich elements correlated with
highly regulated, short-lived mRNAs(23, 29) . We have
identified multiple regions of AU-rich sequence in the 3`-untranslated
region of the IL-8RB gene(25) .)Similarly, the stimulatory effect of G-CSF on chemotactic
responses to IL-8 was more rapid than would be expected from the
transcriptional regulation. These apparent discrepancies may relate to
differences in assay sensitivity or to independent effects of these
stimuli on receptor turnover. Nevertheless, our data suggest that
regulation of the IL-8R after some hours of PMN exposure to G-CSF or
LPS is correlated with the transcriptional regulatory mechanisms
defined here., TNF-, and IL-8(30) , but
also in anti-inflammatory effects leading to decreased PMN
responsiveness such as IL-1RA production (30) and
down-regulation of IL-8R as we have documented here. Perhaps these
immunomodulatory effects of LPS may preferentially direct PMN toward
bacterially derived chemoattractants, such as fMLP. The kinetics and
magnitude of these respective PMN responses may be important
determinants of the outcome of acute neutrophilic inflammation.
), tumor
necrosis factor ; LPS, lipopolysaccharide(s); fMLP,
formylmethionylleucylphenylalanine; G-CSF, granulocyte-colony
stimulating factor; GM-CSF, granulocyte macrophage-colony stimulating
factor; CAT, chloramphenicol acetyltransferase.)
The critical appraisal of the manuscript by Dan Longo
as well as the technical support provided by Karen Blodgett and
Kathleen Bengali are gratefully acknowledged.
©1995 by The American Society for Biochemistry and Molecular Biology, Inc.
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T. Doroshenko, Y. Chaly, V. Savitskiy, O. Maslakova, A. Portyanko, I. Gorudko, and N. N. Voitenok Phagocytosing neutrophils down-regulate the expression of chemokine receptors CXCR1 and CXCR2 Blood, September 18, 2002; 100(7): 2668 - 2671. [Abstract] [Full Text] [PDF]
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B. G. Yipp, G. Andonegui, C. J. Howlett, S. M. Robbins, T. Hartung, M. Ho, and P. Kubes Profound Differences in Leukocyte-Endothelial Cell Responses to Lipopolysaccharide Versus Lipoteichoic Acid J. Immunol., May 1, 2002; 168(9): 4650 - 4658. [Abstract] [Full Text] [PDF]
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H. Nagase, M. Miyamasu, M. Yamaguchi, M. Imanishi, N. H. Tsuno, K. Matsushima, K. Yamamoto, Y. Morita, and K. Hirai Cytokine-mediated regulation of CXCR4 expression in human neutrophils J. Leukoc. Biol., April 1, 2002; 71(4): 711 - 717. [Abstract] [Full Text] [PDF]
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S. S. Cheng, N. W. Lukacs, and S. L. Kunkel Eotaxin/CCL11 Suppresses IL-8/CXCL8 Secretion from Human Dermal Microvascular Endothelial Cells J. Immunol., March 15, 2002; 168(6): 2887 - 2894. [Abstract] [Full Text] [PDF]
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Y.-H. Cui, Y. Le, W. Gong, P. Proost, J. Van Damme, W. J. Murphy, and J. M. Wang Bacterial Lipopolysaccharide Selectively Up-Regulates the Function of the Chemotactic Peptide Receptor Formyl Peptide Receptor 2 in Murine Microglial Cells J. Immunol., January 1, 2002; 168(1): 434 - 442. [Abstract] [Full Text] [PDF]
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S. S. Cheng, J. J. Lai, N. W. Lukacs, and S. L. Kunkel Granulocyte-Macrophage Colony Stimulating Factor Up-Regulates CCR1 in Human Neutrophils J. Immunol., January 15, 2001; 166(2): 1178 - 1184. [Abstract] [Full Text] [PDF]
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S. Yamashiro, J.-M. Wang, D. Yang, W.-H. Gong, H. Kamohara, and T. Yoshimura Expression of CCR6 and CD83 by cytokine-activated human neutrophils Blood, December 1, 2000; 96(12): 3958 - 3963. [Abstract] [Full Text] [PDF]
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C. Murdoch and A. Finn Chemokine receptors and their role in inflammation and infectious diseases Blood, May 15, 2000; 95(10): 3032 - 3043. [Abstract] [Full Text] [PDF]
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R. Bonecchi, F. Facchetti, S. Dusi, W. Luini, D. Lissandrini, M. Simmelink, M. Locati, S. Bernasconi, P. Allavena, E. Brandt, et al. Induction of Functional IL-8 Receptors by IL-4 and IL-13 in Human Monocytes J. Immunol., April 1, 2000; 164(7): 3862 - 3869. [Abstract] [Full Text] [PDF]
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 A. Saccani, S. Saccani, S. Orlando, M. Sironi, S. Bernasconi, P. Ghezzi, A. Mantovani, and A. Sica Redox regulation of chemokine receptor expression PNAS, March 14, 2000; 97(6): 2761 - 2766. [Abstract] [Full Text] [PDF]
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G. Penton-Rol, M. Cota, N. Polentarutti, W. Luini, S. Bernasconi, A. Borsatti, A. Sica, G. J. LaRosa, S. Sozzani, G. Poli, et al. Up-Regulation of CCR2 Chemokine Receptor Expression and Increased Susceptibility to the Multitropic HIV Strain 89.6 in Monocytes Exposed to Glucocorticoid Hormones J. Immunol., September 15, 1999; 163(6): 3524 - 3529. [Abstract] [Full Text] [PDF]
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 W. E. FIBBE, J. F.M. PRUIJT, G. A. VELDERS, G. OPDENAKKER, G. V. KOOYK, C. G. FIGDOR, and R. WILLEMZE Biology of IL-8-Induced Stem Cell Mobilization Ann. N.Y. Acad. Sci., April 30, 1999; 872(1): 71 - 82. [Abstract] [Full Text] [PDF]
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M. H. Khandaker, G. Mitchell, L. Xu, J. D. Andrews, R. Singh, H. Leung, J. Madrenas, S. S.G. Ferguson, R. D. Feldman, and D. J. Kelvin Metalloproteinases Are Involved in Lipopolysaccharide- and Tumor Necrosis Factor-alpha -Mediated Regulation of CXCR1 and CXCR2 Chemokine Receptor Expression Blood, April 1, 1999; 93(7): 2173 - 2185. [Abstract] [Full Text] [PDF]
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