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Laboratories of
*
Host Defenses and
Viral Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892
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Abstract |
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 Top Abstract IntroductionMaterials and MethodsResultsDiscussionReferences |
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and eotaxin (the selective
ligands for CCR1 and CCR3, respectively); and exhibited specific
calcium flux and chemotaxis responses to macrophage-inflammatory
protein-1, eotaxin, and other known CCR1 and CCR3 agonists. CCR3 was
expressed later and at lower levels than CCR1 and could be further
induced by IL-5, whereas IL-5 had little or no effect on CCR1
expression. Consistent with the HIV-1 coreceptor activity of CCR3,
HL-60 clone 15 cells induced with butyric acid and IL-5 fused with HeLa
cells expressing CCR3-tropic HIV-1 envelope glycoproteins, and
fusion was blocked specifically by eotaxin or an anti-CCR3
mAb. These data suggest that CCR1 and CCR3 are markers of late
eosinophil differentiation that are differentially regulated by IL-5 in
this model. |
Introduction |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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(MIP-1) is selective
for CCR1 (2, 3, 5, 7, 8); both receptors are expressed in eosinophils
(6, 7, 8, 9, 15). The importance of CCR3 for eosinophil responses to eotaxin,
RANTES, and MCP-3 has been demonstrated by the blocking effects of an
anti-CCR3 mAb (16); the role of CCR1 in eosinophil responses to
MIP-1 is less well-defined. In addition to their role in leukocyte migration, specific chemokine receptors also act in concert with CD4 as HIV-1 coreceptors, mediating the first step in the viral life cycle, fusion of the viral envelope with the target cell membrane (17, 18, 19, 20, 21, 22). The viral determinant of fusion is the envelope glycoprotein (Env). Different viral strains interact with specific chemokine receptors as determined by sequences in the gp120 component of Env (reviewed in 23 . When CCR3 is expressed in foreign cells, it can support cell fusion reactions mediated by Envs from diverse strains of HIV-1, including those used separately by the major HIV-1 coreceptors CCR5 and CXCR4 (17, 18, 19, 20, 21, 22, 24, 25) (H. Bazan, G. Alkhatib, C. Broder, and E. A. Berger, manuscript in preparation). Moreover, endogenous CCR3 has recently been shown to support HIV-1 infection of microglial cells (24). However, its importance for HIV-1 infection of eosinophils and for HIV-1 pathogenesis has not been demonstrated.
Here we use a cultured HL-60 cell model of eosinophil differentiation to address the ontogeny and regulation of eosinophil chemokine receptor expression. Our results indicate that CCR1 and CCR3 are expressed late during eosinophil differentiation in this model and are differentially regulated by IL-5. Moreover, the data suggest that endogenous eosinophil CCR3 can functionally interact with HIV-1 Envs to facilitate membrane fusion reactions.
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Materials and Methods |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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The promyelocytic cell line HL-60 clone 15 (CRL 1964, American Type Culture Collection, Rockville, MD) was maintained in RPMI 1640 with L-glutamine (Biofluids, Rockville, MD) containing 10% heat-inactivated FCS (HyClone, Logan, UT) and 25 mM N-[2-hydroxyethyl]piperazine-N'-[2-hydroxypropanesulfonic acid] (Sigma Chemical Co., St. Louis, MO), pH 7.6, at 37°C in an atmosphere containing 5% CO2. Cells were induced to differentiate to eosinophil-like cells using 0.5 µM butyric acid (Sigma) as previously described (26, 27). Previously, it has been demonstrated that stimulation with butyric acid for 2 days renders these cells responsive to IL-5 owing to induction of surface IL-5 receptor expression (28). Therefore, in some experiments, 10 ng/ml IL-5 (R&D Systems, Minneapolis, MN) were added to the culture 2 days after addition of butyric acid. After addition of butyric acid, the medium was not replenished, including when IL-5 was added.
Calcium flux assay
Cells were suspended at 1 to 3 x 106/ml in PBS containing 2 µM fura-2/AM (Molecular Probes, Eugene, OR) and incubated for 30 to 60 min at 37°C in the dark. They were then washed twice in HBSS (BioWhittaker, Walkersville, MD) and resuspended in HBSS at 1 x 106 cells/ml. Chemokines were added at indicated times to 1 x 106 cells in a 2-ml volume in a continuously stirred cuvette at 37°C in a Model MS-III fluorimeter (Photon Technology, Inc., South Brunswick, NJ). The relative ratio of fluorescence emitted at 510 nm following sequential excitation at 340 and 380 nm was recorded every 200 ms. Blocking experiments were conducted with the anti-CCR3 mAb 7B11 (generously provided by Charles MacKay), using methods previously described (16).
RNA analysis
Cells were harvested and total RNA prepared using a kit based on a guanidine thiocyanate/phenol extraction method following the manufacturer’s instructions (Stratagene, La Jolla, CA). Isolated RNA (15 µg/sample) was electrophoresed in a 1% agarose gel in 10 mM 3-morpholinopropanesulfonic acid buffer, 5 mM sodium acetate, and 1 mM EDTA at pH 7.0 containing 2% formaldehyde and 10 µg/ml ethidium bromide. The RNA was blotted overnight to Nytran using a Turboblotter apparatus (Schleicher & Schuell, Keene, NH) followed by UV cross-linking. Chemokine receptor probes were labeled using the Random Primer Labeling Kit (Boehringer Mannheim, Indianapolis, IN), and [32P]dCTP, 6000 Ci/mmol (Amersham Corp., Arlington Heights, IL). Northern blots were prehybridized in 50% formamide, 20% dextran sulfate, 5x standard saline-phosphate-EDTA, 0.5% SDS, and 50 µg/ml denatured salmon sperm DNA for 1 h at 37°C. Denatured 32P-labeled probe at 2 x 106 cpm/ml was hybridized to the blot overnight at 37°C. Blots were then rinsed three times in 1x SSC, 0.1% SDS at room temperature, washed at 60°C for 30 to 60 min in 1x SSC, 0.1% SDS, and then exposed to x-ray film. The probes included the complete open reading frame and, in most cases, some untranslated sequence. They are: p4 cDNA, CCR1 (3); clone 3 cDNA, CCR3 (6); PCR-amplified CCR4 open reading frame (A. Sen and J.-L. Gao, unpublished probe); and 8c3 cDNA, CCR5 (13). A 50-mer oligonucleotide probe specific for human ß-actin was used as a control for loading.
Chemokine binding assay
Recombinant human chemokines RANTES, MIP-1, MIP-1ß, MCP-3,
MCP-1, IL-8, and eotaxin were purchased from Peprotech (Princeton, NJ).
125I-labeled human MIP-1, MCP-3, and eotaxin, each
having a sp. act. of 2200 Ci/mmol, were purchased from New England
Nuclear (Boston, MA). Cells were suspended in RPMI 1640 containing 1%
BSA plus azide at 2 x 107/ml. In a 1.5-ml microfuge
tube, increasing concentrations of unlabeled ligand and 0.4 nM
125I-ligand were added to 106 cells in a total
volume of 100 µl. Cells were incubated at room temperature for 1
h with occasional gentle shaking. One milliliter of 10% sucrose in PBS
was then added, and the cells were pelleted in the microfuge tube for 2
min. The supernatants were removed, and gamma emissions from the cell
pellets were counted. Binding data were analyzed using the program
LIGAND.
Chemotaxis
Cells were harvested and washed twice with PBS and then resuspended in serum-free RPMI 1640. Cells (175,000–190,000 per replicate) were loaded in a total volume of 25 µl into the upper compartment of a microchemotaxis chamber (Neuroprobe, Cabin John, MD). Chemoattractants were loaded in a final volume of 31 µl at indicated concentrations into the lower compartment. The two compartments were separated by a polyvinylpyrrolidone-free polycarbonate filter with 5-µm pores. The chemotaxis chamber was incubated at 37°C, 100% humidity, and 5% CO2 for 4 h. The filter was then removed, and the number of cells migrating into the bottom compartment was counted with a hemocytometer. All conditions were tested in triplicate.
Cell fusion assay
HIV Env-mediated cell fusion was quantitated using a vaccinia-based reporter gene activation assay (29). Induced and uninduced HL-60 clone 15 cells were coinfected overnight with recombinant vaccinia viruses vTF7–3 encoding the T7 RNA polymerase (30) and vCB-3 encoding human CD4 (31). The Env from the dual tropic primary HIV-1 isolate 89.6 was cloned into the plasmid vector pSC59 which contains a vaccinia strong early/strong late promoter (H. Bazan, G. Alkhatib, C. Broder, and E. A. Berger, manuscript in preparation; S Chakrabarti and B. Moss, unpublished data). Effector HeLa cells were transfected by lipofection with this plasmid and then infected with recombinant vaccinia virus vCB21R-LacZ containing the ß-galactosidase gene linked to the T7 promoter (30). Cells were then mixed in an HL-60:HeLa ratio of 3:1, in a total volume of 200 µl, and incubated at 37°C for 3 h. Relative cell fusion was recorded as ß-galactosidase activity (OD570 x 1000/min). Blocking experiments were conducted at the start of the coculture by adding either eotaxin or anti-receptor Abs, including the anti-CCR3 mouse mAb 7B11 (16) and a rabbit polyclonal anti-CXCR4 Ab previously described (17). In separate experiments, HeLa cells were coinfected with vCB21R-LacZ, and either vCB-43, encoding Env from the prototypic macrophage-tropic HIV-1 strain Ba-L, or vCB-16, encoding a mutated nonfunctional Env, named Unc, derived from strain IIIB (31, 32, 33). Cells were then mixed with target cells and analyzed as above. Both Ba-L and 89.6 Envs are able to induce cell fusion formation in this assay.
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Results |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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We and others have previously shown that the clone 15 variant of
HL-60 cells can be induced by butyric acid treatment to differentiate
within 2 days into cells having many of the characteristics of
peripheral blood eosinophils, including expression of
eosinophil-specific granule proteins (26, 27). Using Northern blot
analysis, we were unable to detect mRNA for CCR1, -3, -4, or -5 in the
uninduced cells at time zero (Fig. 1
and
data not shown). CCR4 and CCR5 mRNAs remained undetectable for at least
6 days after treatment with butyric acid (not shown). In contrast, CCR1
mRNA was detectable 2 days after addition of butyric acid and increased
progressively over the 5-day period of study (Fig. 1, top). CCR3 mRNA was also induced by butyric acid but
was not detected until 
5 days after addition of butyric acid (Fig. 1, middle). Butyric acid induces expression of IL-5
receptors on HL-60 clone 15 cells, and the cells then proliferate in
response to IL-5 (28). We observed that the level of expression of CCR3
was markedly increased by the addition of IL-5 to the butyric
acid-treated culture, the concentration previously shown to be optimal
for proliferation of these cells. In contrast to its effects on steady
state CCR3 mRNA, IL-5 had little effect on the amount of CCR1 mRNA
detected (Fig. 1).
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To test whether induction of CCR1 and CCR3 mRNA correlated with
induction of binding sites for known CCR1 and CCR3 ligands, we
conducted radiolabeled chemokine binding assays. Specific binding of
125I-MIP-1, 125I-MCP-3, or
125I-eotaxin was not detected on uninduced HL-60 cells when
0.4 nM radioligand was used as a probe (not shown). Three days after
butyric acid was added, specific binding of 125I-MIP-1
and 125I-MCP-3, but not 125I-eotaxin, was
detected (Fig. 2A).
125I-MIP-1 binding was strongly inhibited in the
presence of 100 nM unlabeled MIP-1 (95% inhibition), RANTES (68%),
MCP-3 (90%), MIP-1ß (75%), and MCP-1 (65%), but not by the CXC
chemokine IL-8 (0%). Eotaxin reduced binding by only 18%, indicating
that the great majority of MIP-1- and eotaxin-binding sites are
distinct. Although statistically significant, the functional
significance of this small amount of cross-competition is unclear,
especially since we did not observe it later in the course of cell
differentiation, and since direct eotaxin binding and signaling was not
observed (see below). The competition profile for the
125I-MCP-3-labeled site was similar, although not
identical. The specificity of competition for the MIP-1 binding site
is consistent with that established previously for recombinant
CCR1 (2).
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and
125I-MCP-3 were similar to the profiles observed for cells
treated with butyric acid alone (Fig. 2
B). However,
under these conditions, specific binding of 125I-eotaxin
was also detected (Fig. 2B). Eotaxin binding was
strongly inhibited by 100 nM unlabeled eotaxin (75% inhibition) and
MCP-3 (53%) but was affected only weakly by RANTES (30%), MCP-1
(20%), and MIP-1ß (15%) and not at all by IL-8 and MIP-1. The
specificity of competition for the eotaxin-binding site is consistent
with that established previously for primary eosinophils and
recombinant CCR3 (8, 15). Thus, at this time point eotaxin and MIP-1
appear to bind to separate sites on HL-60 clone 15-derived eosinophils,
and MIP-1ß, MCP-1, MCP-3, and RANTES appear to share binding
determinants with both. The induced cells expressed 23,000
eotaxin-binding sites/cell with a Kd of 2.6 nM.
Unlabeled MIP-1, RANTES, and MCP-3 competed for the
125I-MIP-1-labeled site on induced cells with
IC50s 5, 20, and 50 nM, respectively (Fig. 3B). The cells
expressed 650,000 MIP-1 binding sites/cell with a
Kd of 42 nM.
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To test whether the chemokine-binding sites on HL-60 clone
15-derived eosinophils were functional, we first monitored changes in
[Ca2+]i in response to stimulation with
chemokines. A rapid, transient calcium flux is typically observed when
leukocytes are stimulated with chemokines, and it serves as a
convenient measure of chemokine receptor activation that can be
followed in real time in fura-2-loaded cells. Uninduced cells and cells
treated with IL-5 alone responded negligibly to MIP-1, RANTES,
MCP-3, FMLP, eotaxin, MIP-1ß, MCP-1, or IL-8. In contrast, the cells
responded well to ATP which activates purinergic receptors in myeloid
cells (Fig. 4 and data not shown). HL-60
clone 15 cells cultured in the presence of butyric acid acquired
responsiveness to MIP-1, RANTES, MCP-3, eotaxin, and FMLP (Fig. 4).
The cells did not respond to MIP-1ß, consistent with absence of
detectable mRNA for CCR5, the only known MIP-1ß receptor. The cells
also did not respond to MCP-1 or IL-8, which suggests that the IL-8
receptors CXCR1 and CXCR2, and the MCP-1 receptor CCR2 are also are not
expressed in these cells, although we did not specifically probe for
the corresponding mRNAs.
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. The responses to RANTES, MCP-3, and FMLP were similar in
magnitude to MIP-1 and increased rapidly by day 2 and peaked on day
4 after addition of butyric acid (Figs. 4 and 5, A and
B, and data not shown). In contrast, the response to eotaxin
was never observed on day 2 after addition of butyric acid but was
consistently observed by day 4 (Fig. 5).
Adding IL-5 to butyric acid-treated cells markedly enhanced
responsiveness to eotaxin, beginning on day 4 and peaking on day 6, but
had little effect on responsiveness to MIP-1, RANTES, MCP-3, and
FMLP (Fig. 5 and data not shown). The 50% effective concentration
(EC50) of the calcium flux response to eotaxin for HL-60
clone 15 cells induced with butyric acid and IL-5 was 5 nM, a value
similar to the KD for eotaxin binding (Fig. 6). Eotaxin has similar potency for
calcium flux at recombinant CCR3 expressed in HEK 293 cells (7). The
anti-CCR3 mAb 7B11 completely blocked the calcium flux response to
eotaxin in HL-60 clone 15 cells, but had no effect on the response to
MIP-1 (Fig. 7). It also specifically
blocked the eotaxin-induced calcium flux response in mouse pre-B cells
transfected with recombinant CCR3 (J. E. Pease and P. M.
Murphy, unpublished data), confirming a previous report (16).
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We next tested the ability of CCR1 and CCR3 ligands to induce
chemotaxis in HL-60 clone 15-derived eosinophils (Fig. 8). Uninduced HL-60 clone 15 cells did
not move in response to either MIP-1 or eotaxin (data not shown).
Cells treated with butyric acid for 3 days moved weakly in response to
MIP-1 with an optimal concentration of 5 nM, whereas the same cells
did not move in response to eotaxin at any concentration tested (range,
1 to 500 nM). In contrast, cells treated under optimal conditions for
expression of CCR1 and CCR3 (butyric acid plus IL-5) exhibited robust
chemotactic responses to both eotaxin and MIP-1. As would be
predicted from results of the calcium flux assay, MIP-1 was a much
more effective agonist for chemotaxis. A typical bell-shaped
dose-response curve was observed for both chemokines, with
EC50s of 5 and 20 nM for MIP-1 and eotaxin,
respectively. Consistent with results from the calcium flux assay,
RANTES and MCP-3 (but not MIP-1ß, MCP-1, and IL-8) were
able to chemoattract induced HL-60 clone 15 cells (data not shown).
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As a fourth test of CCR3 induction in butyric acid and
IL-5-treated HL-60 clone 15 cells, we conducted HIV-1 Env-dependent
cell fusion assays (Fig. 9). We and
others have previously shown that CCR3 facilitates membrane fusion and
cell entry for certain HIV-1 strains, including the prototypic dual
tropic primary HIV-1 strain 89.6 and the prototypic macrophage-tropic
strain Ba-L (21, 22, 24, 25) (H. Bazan, G. Alkhatib, C. Broder, and
E. A. Berger, manuscript in preparation). Uninduced HL-60 cells
showed low levels of fusion with cells expressing either Ba-L or 89.6
Env, that were markedly increased after treatment with butyric acid and
IL-5 (Fig. 9). Addition of eotaxin to the coculture suppressed fusion
in a dose-dependent manner, with an EC50 of 10 nM
for the 89.6 Env and a threshold of 100 nM for the Ba-L Env (Fig. 9, A and D). The reason for the difference in
blocking potency by eotaxin for the two Envs is not known but may
reflect differences in levels of Env expression, differences in the
CCR3 interaction sites for these Envs, and/or differential usage of
other unknown coreceptors. RANTES and MCP-3, which bind to CCR3 with
30- and 100-fold lower affinity than eotaxin, respectively (8), were
unable to block Ba-L Env-dependent fusion of induced clone 15 cells
(Fig. 9D), consistent with previous results using
National Institutes of Health 3T3 cells expressing recombinant
CCR3 (25).
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50%, whereas control Abs had a negligible effect (Fig. 9, B and C). Consistent with previous reports
of the ability of the 89.6 Env to interact with both recombinant CXCR4
and CCR3, we observed increased inhibition of fusion when both Abs were
added together vs either added alone (Fig. 9C). |
Discussion |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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The MIP-1-binding site on HL-60 clone 15-derived eosinophils is
similar to that reported previously for HL-60 clone 7-derived
eosinophils (34). The eotaxin-binding site on clone 15 cells is similar
to that described on primary eosinophils (15). Taken together, the
results are consistent with the model of chemokine binding established
from studies of CCR1 and CCR3 in heterologous transfected cells (2, 8).
Specifically, MIP-1 and eotaxin bind to separate noninteracting
sites, both of which overlap with binding determinants with other CC
chemokines including RANTES and MCP-3.
Although the relative levels of expression of CCR1 and CCR3 in
differentiated HL-60 clone 15 cells are reciprocal to those found in
primary eosinophils (6, 8, 9, 15), the cell line is a useful surrogate
for eosinophils in studies requiring large numbers of cells or
observations over time in vitro. In this regard, our study
provides new information about factors that regulate CCR1 and CCR3
expression, specifically butyric acid and IL-5. CCR1 and CCR3 mRNA
levels accumulate in the HL-60 clone 15 cell line treated with butyric
acid, which also induces the cells to terminally differentiate. Since
the uninduced cells are arrested at the promyelocytic stage of
myelopoiesis, CCR1 and CCR3 appear to be markers of mature eosinophils
and not of precursor cells, although additional studies with primary
bone marrow-derived cells will be needed to confirm this. CCR1 mRNA
appeared much sooner than CCR3 mRNA and accumulated to much higher
levels after butyric acid induction. This correlated well with the
ontogeny and magnitude of cell responses to MIP-1 and eotaxin, the
selective agonists for CCR1 and CCR3, respectively. CCR3 is expressed
later in the HL-60 model of eosinophil maturation than other eosinophil
markers such as eosinophil-derived neurotoxin and eosinophil cationic
protein (26) and appears to represent one of the final steps in the
differentiation process.
Our study also provides the first evidence that IL-5 up-regulates CCR3 expression in differentiating eosinophils and is consistent with the extremely high levels of mouse CCR3 mRNA reported in IL-5 transgenic mice (35). Butyric acid is thought to activate genes by blocking the action of deacetylases, thereby inhibiting histone binding to DNA due to increased histone acetylation (36). Thus, butyric acid may be acting at the chromosomal level to induce CCR1 and CCR3 expression. Although CCR1 expression was not affected by IL-5 in HL-60 clone 15-derived eosinophils, it is up-regulated by IL-2 treatment of T lymphocytes, as is CCR2 (37). IL-5 induction of CCR3 expression could potentially explain why eosinophil recruitment is enhanced at sites injected with IL-5 (38).
Previously, CCR3 was shown to function as an HIV-1 coreceptor (21, 22, 25, 39) (H. Bazan, G. Alkhatib, C. Broder, and E. A. Berger, manuscript in preparation) that facilitates infection of microglial cells in vitro (25), suggesting a potential role for CCR3 in central nervous system infection by HIV-1. Our results suggest that CCR3 could also function as an HIV-1 coreceptor in maturing eosinophils. This is relevant since HIV-1 has been detected in bone marrow eosinophils from certain HIV-1-infected individuals (40, 41); however, the role of eosinophil infection in HIV-1 pathogenesis is not known.
In addition to increasing HIV-1 coreceptor expression on individual cells, IL-5 is an important eosinophilopoietin and, in particular, regulates the hypereosinophilia associated with helminth infections (42). In underdeveloped world regions where helminth infections are prevalent, these separate effects of IL-5 could conspire to facilitate HIV-1 transmission and to accelerate immune system deterioration and progression to AIDS in infected individuals by providing the virus with a greatly expanded target area.
Although RANTES and MCP-3 are ligands for CCR3, neither was an inhibitor of HIV-1 Env-dependent fusion. This is consistent with their relatively lower affinity and potency compared with eotaxin established by studies of CCR3 in transfected pre-B cell lymphoma cells (8) and suggests that development of CCR3-directed antagonists of HIV-1 entry might be more successful starting from an eotaxin prototype than from RANTES or MCP-3.
In summary, the HL-60 clone 15 cell line can be used as a model of CCR1 and CCR3 regulation and CCR3 interactions with HIV-1 Envs. Not only can this cell line be used to follow expression of these receptors during development but it also can be used to study receptor antagonists as they become available both for chemokines and for HIV-1. The cell line should also be useful for detailed studies of gene regulation and signal transduction for CCR1 and CCR3.
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Footnotes |
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2 Address correspondence and reprint requests to Dr. Philip M. Murphy, Building 10, Room 11N113, National Institutes of Health, Bethesda, MD 20892. E-mail address:
3 Abbreviations used in this paper: CCR, CC chemokine receptor; MIP-1, macrophage inflammatory protein-1; MCP, monocyte chemoattractant protein; Env, envelope glycoprotein of HIV-1; EC50, 50% effective concentration.
Received for publication March 20, 1997. Accepted for publication October 9, 1997.
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References |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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/RANTES receptor. J. Exp. Med. 177:1421., MIP-1ß, and RANTES. J. Leukocyte Biol. 60:147.[Abstract]
. J. Biol. Chem. 271:17161., MIP-1ß receptor as a fusion cofactor for macrophage-tropic HIV-1. Science 272:1955.[Abstract]
upon differentiation of an eosinophilic HL-60 cell line. J. Immunol. 152:4055.[Abstract]
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N. Zimmermann, B. L. Daugherty, J. M. Stark, and M. E. Rothenberg Molecular Analysis of CCR-3 Events in Eosinophilic Cells J. Immunol., January 15, 2000; 164(2): 1055 - 1064. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. M. Burns, G. K. Lewis, and A. L. DeVico Soluble complexes of regulated upon activation, normal T cells expressed and secreted (RANTES) and glycosaminoglycans suppress HIV-1 infection but do not induce Ca2+ signaling PNAS, December 7, 1999; 96(25): 14499 - 14504. [Abstract] [Full Text] [PDF]
|
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 |
|
 K. S. C. Weber, P. J. Nelson, H.-J. Grone, and C. Weber Expression of CCR2 by Endothelial Cells : Implications for MCP-1 Mediated Wound Injury Repair and In Vivo Inflammatory Activation of Endothelium Arterioscler. Thromb. Vasc. Biol., September 1, 1999; 19(9): 2085 - 2093. [Abstract] [Full Text] [PDF]
|
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|
 M. A. Giembycz and M. A. Lindsay Pharmacology of the Eosinophil Pharmacol. Rev., June 1, 1999; 51(2): 213 - 340. [Abstract] [Full Text] [PDF]
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0.005 by ANOVA) for all unlabeled
chemokines tested except for IL-8 (all panels) and eotaxin inhibition
of 125I-MIP-1
), RANTES (
) or MCP-3
(
). Insets at the upper right of each panel show
Scatchard analysis of the binding data indicating a
Kd of 42 nM for MIP-1
) or after addition of butyric acid (BA) 0.5 µM at
time 0 (
), IL-5 10 ng/ml alone at day 2 (
