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*
Division of Viral Immunology, Center for AIDS Research, Kumamoto University, and
Chemo-Sero Therapeutic Research Institute, Kyokushi, Kumamoto, Japan; and
AIDS Clinical Center, International Medical Center of Japan, Tokyo, Japan
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Abstract |
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 Top Abstract IntroductionMaterials and MethodsResultsDiscussionReferences |
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CCR5+CCR7+CCR5+CCR7-.
CCR5+CD8+ T cells effectively migrated in
response to RANTES, suggesting that CCR5 plays a critical role in the
migration of Ag-specific effector and differentiated memory
CD8+ T cells to inflammatory tissues and secondary lymphoid
tissues. This is in contrast to CCR7, which functions as a homing
receptor in migration of naive and memory CD8+ T cells to
secondary lymphoid tissues. |
Introduction |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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CCR5 is a receptor for 
-chemokines RANTES and
macrophage-inflammatory protein
(MIP)3-1
and -1,
and acts as a coreceptor for HIV-1 entry (3, 4, 5). The
expression of this receptor has been well characterized in
CD4+ T cells: in peripheral blood CCR5 is
expressed only on Th1 CD4+ T cells
(6), while in thymus it is not expressed on
CD3- immature thymocytes but is expressed at low
levels on
CD3+CD4-CD8+
and
CD3+CD4+CD8-
thymocytes (7). A few studies have examined CCR5
expression in CD8+ T cells. In mice, Tc1 effector
CD8+ T cells, which produce type 1 cytokines,
predominantly express CCR5 mRNA, while Tc2 effector
CD8+ T cells, which produce type 2 cytokines,
express high levels of CXCR4 mRNA but not CCR5 mRNA
(8, 9, 10). In humans, previous studies showed that a subset
of CD8+ T cells expresses CCR5
(11, 12, 13, 14). However, the expression and function of this
chemokine receptor on human CD8+ T cells remain
unclear.
CCR7 is another chemokine receptor and functions as a homing
receptor in migration of naive and memory CD8+ T
cells to secondary lymphoid tissues. A role for CCR7 in
CD8+ T cells has been demonstrated through the
identification of
CCR7+CD45RA+ naive
CD8+ T cells and
CCR7+CD45RA- and
CCR7-CD45RA- memory
CD8+ T cells (15). Furthermore, a
recent study suggested the following differentiation lineage for
Ag-specific CD8+ T cells:
CCR7+CD45RA+CCR7+CD45RA-CCR7-CD45RA-CCR7-CD45RA+
(16). This study suggested that
CCR7+CD45RA+CD8+
and
CCR7+CD45RA-CD8+
T cells are precursor memory T cells while
CCR7-CD45RA-CD8+
and
CCR7-CD45RA+CD8+
T cells are preterminally differentiated and terminally differentiated
cells, respectively.
In the present study, we examined in detail the surface expression of CCR5 and CCR7 on CD28CD45RA subpopulations in human total CD8+ T cells and Ag-specific CD8+ T cells. In addition, the function of CCR5 on human CD8+ T cells was also analyzed. We suggest a different role for CCR5 and CCR7 in the Th1 immune response.
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Materials and Methods |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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Anti-human CCR7 mAb (2H4), FITC-conjugated anti-human CCR5
mAb (2D7), PE- and allophycocyanin-conjugated anti-human
CD28 mAb, CyChrome-conjugated anti-human CD45RA mAb,
PerCP-conjugated anti-human CD8 mAb, and FITC- and
biotin-conjugated rat anti-mouse IgM mAb were purchased from BD
PharMingen (San Diego, CA). Allophycocyanin-conjugated anti-human
CD8 mAb was purchased from Beckman Coulter (Fullerton, CA).
PE-conjugated extravidin was purchased from Sigma Chemical (Irvine,
CA). The anti-human CCR5 mAb (45531.111) used for the lymphocyte
chemotaxis assay was purchased from DAKO (Kyoto, Japan). Recombinant
human RANTES/CCL5 and MIP-1 were purchased from R&D Systems
(Minneapolis, MN).
Blood samples
PBMCs from 13 individuals with chronic HIV-1 infection were analyzed in this study. Blood samples were collected with oral informed consent from HLA-A11+, HLA-A24+, and/or HLA-B35+ HIV-1-seropositive individuals at the International Medical Center of Japan (Tokyo, Japan). All these individuals belonged to the clinical stage of asymptomatic carrier or AIDS-related complex. PBMCs from 11 HIV-1-seronegative healthy subjects were also analyzed in this study. Two HLA-A*0201 samples were used for analysis of human CMV (HCMV)-specific CD8+ T cells. Blood samples were also collected with oral informed consent.
Synthetic peptides
Sixteen CTL epitope peptides were synthesized using an automated multiple peptide synthesizer (Shimadzu Model PSSM-8; Shimadzu, Kyoto, Japan). These peptides were as follows: an HLA-A*0201-restricted HCMV epitope (HCMV-pp65 495–503 NLVPMVATV) (17), an HLA-B*3501-restricted hepatitis C virus (HCV) epitope (HCV-NS3 1359–1367 HPNIEEVAL) (18), two HLA-A*1101-restricted HIV epitopes (HIV-Pol 313–321 AIFQSSMTK, and -Nef 84–92 AVDLSHFLK) (19, 20), five HLA-A*2402-restricted HIV epitopes (HIV-Nef 138–147 RYPLTFGWCF, -Nef 138–147-2F RFPLTFGWCF, -env 584–592 RYLRDQQLL, -env 679–687 WYIKIFIMI, and -Gag 28–36 KYKLKHIVW) (21, 22), and five HLA-B*3501-restricted HIV epitopes (HIV-Pol 273–282 VPLDKDFRKY, -Nef 75–85 RPQVPLRPMTY, -Pol 448–456 IPLTEEAEL, -Pol 587–595 EPIVGAETF, and -env 77–85 DPNPQEVVL) (23) and two HLA-A*1101-restricted EBV epitopes (EBV-3B 416–424: IVTDFSVIK, and EBV-3B 399–408: AVFDRKSDAK) (24). All peptides were verified to be >90% pure by mass spectrometry and HPLC.
Production of HLA class I/peptide tetramers
HLA-A*0201, HLA-A*1101, HLA-A*2402, and HLA-B*3501/peptide
tetrameric complexes were prepared as previously described (25, 26). Briefly, recombinant human 2
microglobulin and recombinant HLA class I derivatives (COOH termini of
HLA class I molecules with truncated transmembrane and cytoplasmic
domains, and with a sequence containing the BirA enzymatic
biotinylation site) were purified from Escherichia coli
cells transformed with the relevant expression plasmid. Monomeric
complexes were generated by in vitro refolding of human
2 microglobulin, the appropriate HLA class I
derivative, and an HIV-1-specific, EBV-specific, or HCMV-specific
epitope peptide. The resultant 45-kDa complexes were separated by gel
filtration using a Superdex G75 column (Amersham Pharmacia Biotech,
Uppsala, Sweden) and then biotinylated enzymatically with BirA enzyme
(Avidity, Denver, CO). The biotin-binding complexes were separated by
gel filtration using a Superdex G75 column followed by anion exchange
using a Mono Q column (Amersham Pharmacia Biotech). HLA class I/peptide
tetramers were generated by mixing the monomer complexes with
PE-conjugated extravidin (Sigma Chemical) or allophycocyanin-conjugated
streptavidin (BD PharMingen) at a molar ratio of 4:1.
CTL clones
Three HIV-specific CTL clones for HIV-Pol 448–456, HIV-Nef 75–85, and HIV-env 679–687 (HIV-B35-SF2–24-55, HIV-B35-SF2–6-101, HIV-A24-SF2-env 679–687-3, respectively) and an HCV-specific CTL clone for HCV-NS3 1359–1367 (HCV-B35–38-20) were generated previously (18, 21, 23).
Flow cytometry analyses
Cryopreserved PBMCs from HIV-1-seropositive individuals and healthy subjects were stained with mAbs and/or tetramers in various combinations. PBMCs from HIV-1-seronegative and -seropositive individuals were stained with FITC-conjugated anti-CCR5 mAb and PerCP-conjugated anti-CD8 mAb, with those from HIV-1-seropositive individuals further stained with allophycocyanin- or PE-conjugated tetramer(s). Incubation with tetramer(s) and subsequent washing were performed at 37°C to avoid nonspecific binding to CD8+ T cells, as previously described (27). For four-color flow cytometry, PBMCs isolated from healthy individuals were stained with FITC-conjugated anti-CCR5, PE-conjugated anti-CD28, CyChrome-conjugated anti-CD45RA, and allophycocyanin-conjugated anti-CD8. Similarly, after CD8+ T cells were purified from PBMCs of HIV-1-seronegative and -seropositive individuals using the MACS system (Miltenyi Biotec, Bergisch Gladbach, Germany), these cells were stained with FITC-conjugated anti-CCR5 mAb, PE-conjugated anti-CD28 mAb, CyChrome-conjugated anti-CD45RA mAb, and allophycocyanin-conjugated tetramer(s) for HCMV and HIV-1, respectively. CTL clones were stained with FITC-conjugated anti-CCR5 mAb and PerCP-conjugated anti-CD8 mAb. To investigate CCR7 expression on CD8+ T cells and Ag-specific CD8+ T cells, purified CD8+ T cells from HCMV-seropositive healthy individuals were stained with anti-CCR7 mAb followed by FITC-conjugated rat anti-mouse IgM mAb, PE-conjugated anti-CD28 mAb, CyChrome-conjugated anti-CD45RA mAb, and allophycocyanin-conjugated tetramer for HCMV. To investigate coexpression of CCR7 and CCR5 on CD8+ T cells, purified CD8+ T cells from HCMV-seropositive healthy individuals were stained with anti-CCR7 mAb followed by biotin-conjugated rat anti-mouse IgM mAb and then PE-conjugated extravidin, FITC-conjugated anti-CCR5 mAb, CyChrome-conjugated anti-CD45RA mAb, and allophycocyanin-conjugated anti-CD28 mAb. Cells were washed three times with PBS supplemented with 10% FCS and then resuspended with PBS containing 2% paraformaldehyde. Samples were analyzed by FACSCalibur with CellQuest software (BD Biosciences, San Jose, CA).
CTL assay
CTL activity was measured by a standard 51Cr release assay as follows. The HLA-A,-B defective cell line C1R as well as the C1R transfectants C1R-A*1101 and C1R-A*0201 (5 x 105 cells) were incubated for 60 min with 3.7 M Bq Na251CrO4 in saline and then washed three times with RPMI 1640 medium containing 10% newborn calf serum. Labeled target cells (2 x 103/well) were added into U-bottom 96-well microtiter plates with 10 µM HCMV (HCMV-pp65 495–503) or two EBV (EBV-3B 416–424 and EBV 3B 399–408) peptides. After incubation for 1 h, CD8+ T cells purified from PBMC of EBV-seropositive, HLA-A11+ individuals by anti-CD8 mAb-coated beads or CD28-CD45RA-CD8+ and CD28-CD45RA+CD8+ T cells purified from PBMC of HCMV-seropositive, HLA-A2+ individual by a cell sorter (FACSVantage SE; BD Biosciences) were added at various effector (tetramer+CD8+ T cells):target ratios. The mixtures were incubated for 6 h at 37°C and then the supernatants were collected and analyzed with a gamma counter. Spontaneous 51Cr release was determined by measuring the cpm in supernatants from wells containing only target cells (cpm spn). Maximum 51Cr release was determined by measuring the cpm in supernatants from wells containing target cells in the presence of 2.5% Triton X-100 (cpm max). Specific lysis = ((cpm exp - cpm spn)/(cpm max - cpm spn)) x 100, where cpm exp is the cpm in supernatants of wells containing both target and effector cells.
Lymphocyte chemotaxis assay
The lymphocyte chemotaxis assay was performed using 96-well microchemotaxis chambers (NeuroProbe, Gaithersburg, MD) as previously described (28). The MACS magnetic cell separation system was used to isolate CD8+ T cells from fresh PBMCs taken from healthy individuals. More than 98% of the purified cells were CD8+ T cells. The purified CD8+ T cells (5 x 105 cells/well) were placed over the filter (5-µm pore diameter) in the upper wells of the chamber. Recombinant human RANTES/CCL5 and an anti-human CCR5 mAb were diluted with RPMI 1640 and then applied to either the upper or lower wells of the chamber. After incubation at 37°C for 2 h, cells remaining above the filter (i.e., cells that did not migrate) were removed by washing with PBS containing 2 mM EDTA and wiping. Cells that had migrated to the lower well of the chamber and below the filter were collected by centrifugation. These cells were counted using a hemocytometer and then stained with PE-conjugated anti-CD28 mAb and CyChrome-conjugated anti-CD45RA mAb for flow cytometry analysis. The percentage of migrated cells with each CD28/CD45RA phenotype was determined as follows: the total number of cells with each phenotype added to the upper well was calculated from total number of cells added to the upper well and the percentage of each phenotype determined by FACS analysis. The migrated cell number of each phenotype was calculated from both the total number of cells migrated to the lower well and the percentage of each phenotype. The percentage of the migrated cells with each phenotype = (migrated cell number with each phenotype/total cell number added to the upper wells with each phenotype) x 100.
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Results |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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We used flow cytometry to investigate the surface expression
of CCR5 on total CD8+ T cells in PBMCs isolated
from 11 healthy individuals. A significant number of total
CD8+ T cells expressed CCR5 (Fig. 1
A). Expression varied among
individuals, with the percentage of
CCR5+CD8+ T cells in the
total CD8+ T cell population ranging from 11.9 to
34.9% (mean, 24.7 ± 8.1%; Fig. 1C). These results
indicate that a subset of CD8+ T cells
expresses CCR5.
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, A and
C), with the percentage of these cells in the total
CD8+ T cell population ranging from 15.1 to
77.3% (mean, 43.1 ± 18.3%; Fig. 1C). As the number
of HIV-1-specific CD8+ T cells is thought to
increase in HIV-1-infected individuals, these results suggest that most
HIV-1-specific CD8+ T cells express CCR5.
Therefore, we investigated the surface expression of CCR5 on
HIV-1-specific CD8+ T cells in PBMCs isolated
from HIV-1-seropositive individuals using HLA-A*2402, HLA-A*1101,
and/or HLA-B*3501 tetramers. Results from a representative individual
(KI-003) are shown in Fig. 1B. HIV-1-specific
CD8+ T cells were detected by the three
HLA-B*3501 tetramers in PBMCs from KI-003 (Fig. 1B,
left panel). These
tetramer+CD8+ T cells
expressed CCR5 (Fig. 1B, right panel). Analysis
of PBMCs from 11 HIV-1-infected individuals showed that the majority of
HIV-1-specific CD8+ T cells expressed CCR5 (mean,
63.7 ± 17.0%; Fig. 1C). CCR5 expression was similarly
observed on HCMV-specific CD8+ T cells (data not
shown). CCR5 was also expressed on HIV-1-specific and HCV-specific
CD8+ CTL clones (Fig. 2), further confirming that Ag-specific
CD8+ T cells express CCR5.
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A previous study showed that CD27+CD45RA+CD8+ T cells have neither perforin nor cytolytic activity while CD27+CD45RA-CD8+, CD27-CD45RA-CD8+, and CD27-CD45RA+CD8+ T cells have a low, medium, and high level of perforin, respectively, and have cytolytic activity in proportion to perforin expression (29). This study also showed that the latter three types of CD8+ T cells can effectively produce cytokines. Similarly, our previous study of CD28, CD45RA, and perforin expression in CD8+ T cells showed that CD28+CD45RA+CD8+ T cells have no perforin while CD28+CD45RA-CD8+, CD28-CD45RA-CD8+, and CD28-CD45RA+CD8+ T cells have a low, medium, and high level of perforin, respectively (25). Therefore, we investigated the cytolytic activity of these CD28CD45RA subsets. HCMV-specific CD28-CD45RA-CD8+ and CD28-CD45RA+CD8+ T cells effectively lysed target cells pulsed with HCMV epitope peptide while EBV-specific CD28+CD45RA-CD8+ T cells failed to kill target cells pulsed with EBV epitope peptide (data not shown). Thus, naive, memory, and effector CD8+ T cells can be discriminated by their different surface expression of CD28(CD27) and CD45RA (CD28+(CD27+)CD45RA+, naive cells; CD28+(CD27+)CD45RA-, memory cells; CD28-(CD27-)CD45RA-, memory/effector cells; and CD28-(CD27-)CD45RA+, effector cells) (25, 29, 30).
The observation that the level of CCR5 expression on
CD8+ T cells varied among individuals suggests
that CCR5 may be expressed at different levels on naive, memory, and/or
effector CD8+ T cells. Therefore, we examined
CCR5 expression in these four CD8+ T cell
populations. PBMCs from one healthy individual were analyzed by
four-color flow cytometry analysis using four mAbs (anti-CD8,
anti-CD28, anti-CD45RA, and anti-CCR5 mAbs).
CD28-CD45RA-CD8+
and
CD28-CD45RA+CD8+
T cells expressed moderate levels of CCR5 while
CD28+CD45RA-CD8+
T cells expressed high levels of CCR5. In contrast,
CD28+CD45RA+CD8+
T cells did not express CCR5 (Fig. 3).
The same results were observed in CD8+ T cells
from eight HIV-1-seronegative healthy individuals (Table I).
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). Thus, in both HIV-1-seropositive and
HCMV-seropositive individuals, CCR5 expression increases during
differentiation of naive CD8+ T cells to memory
CD8+ T cells and then decreases during
differentiation to effector CD8+ T cells.
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Recent studies demonstrated that CCR7 is expressed on naive
(CD27+CD45RA+) and memory
(CD27+CD45RA-)
CD8+ T cells (30). We analyzed CCR7
expression on the different CD8+ T cell
populations from three healthy individuals. Results from a
representative individual are shown in Fig. 5A.
CD28+CD45RA+CD8+
T cells and 
50% of
CD28+CD45RA-CD8+
T cells express CCR7 while
CD28-CD45RA-CD8+
and
CD28-CD45RA+CD8+
T cells did not express CCR7. Thus, we confirmed that naive and memory
CD8+ T cells express CCR7. To clarify whether
Ag-specific memory CD8+ T cells express CCR7, we
analyzed CCR7 expression on HCMV-specific CD8+ T
cells isolated from two HCMV-seropositive individuals. These results
showed that in HCMV-specific CD8+ T cells,
CD28-CD45RA-CD8+
and
CD28-CD45RA+CD8+
T cells did not express CCR7 while a small population of
CD28+CD45RA-CD8+
T cells did express CCR7 (Fig. 5B), suggesting that
HCMV-specific memory CD8+ T cells may be well
differentiated.
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50%
expressed CCR7, suggesting that the memory CD8+ T
cell population has at least two different subsets. Therefore, we
further analyzed expression of CCR5 and CCR7 on
CD28+CD45RA-CD8+
T cells. Results from three individuals demonstrated the existence of
three subsets of
CD28+CD45RA- memory
CD8+ T cells:
CCR5+CCR7-,
CCR5-CCR7+, and
CCR5+CCR7+ (Fig. 6). The expression level of CCR5 in the
CCR5+CCR7- subset was
higher than that in the
CCR5+CCR7+ subset, while
the expression level of CCR7 in the
CCR5-CCR7+ subset was
higher than that in the
CCR5+CCR7+ subset. CCR5 was
expressed on
CD28-CD45RA-CD8+
T cells but not on
CD28+CD45RA+CD8+
T cells, while CCR7 was expressed on
CD28+CD45RA+CD8+
T cells but not on
CD28-CD45RA-CD8+
T cells. These findings strongly suggest the following differential
lineage in
CD28+CD45RA- memory
CD8+ T cells:
CCR5-CCR7+CCR5+CCR7+CCR5+CCR7-.
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Migration of virus-specific memory and effector
CD8+ T cells to regions of viral replication
seems to be one of the most important events in virus eradication. As
CCR5 is a receptor for the -chemokines RANTES, MIP-1, and
MIP-1, CCR5+CD8+ T cells
could be expected to actively migrate to inflammatory regions where
these chemokines are produced. To clarify the function of CCR5 in
CD8+ T cells, we investigated RANTES-mediated
migration of CCR5+CD8+ T
cells. CD8+ T cells were isolated from PBMCs of a
healthy individual and then added to the upper wells of a 96-well
microchemotaxis chamber. The CD28/CD45RA phenotype of cells that
migrated to the bottom well of the chambers was determined by staining
with anti-CD28 mAb and anti-CD45RA mAb followed by flow
cytometry. An effect of RANTES on CD8+ T cell
migration was observed for
CD28+CD45RA-,
CD28-CD45RA-, and
CD28-CD45RA+ populations
in a dose-dependent fashion (Fig. 7A). This effect was stronger
on the CD28-CD45RA+
population than on the other two populations. These results were
confirmed by three independent experiments. The effect of RANTES on
these three populations was abrogated by an anti-CCR5 mAb (Fig.
7B). These three populations did not express CCR1, another
receptor for RANTES (data not shown). MIP-1 had a similar effect on
these three populations (data not shown). MIP-1 also had a weak
effect on the CD28+CD45RA+
population (data not shown), suggesting that this population may
express another receptor for MIP-1. These results together suggest
that memory and effector CD8+ T cells can
actively migrate in response to the -chemokines RANTES and
MIP-1.
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Discussion |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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Expression of CCR5 on CD8+ T cells is thought to
be important in Th1 immune responses associated with -chemokines.
Though previous studies showed that some CD8+ T
cell populations express CCR5 (11, 12, 13, 14), in this study we
demonstrated that CCR5 expression is restricted to
CD28+CD45RA-CD8+,
CD28-CD45RA-CD8+,
and
CD28-CD45RA+CD8+
T cell populations, i.e., memory and effector
CD8+ T cells but not naive
CD8+ T cells express CCR5.
CD8+ T cells have been shown to express the
chemokine receptor CCR7 (15). A previous study suggested
the following differentiation lineage for Ag-specific
CD8+ T cells:
CCR7+CD45RA+CCR7+CD45RA-CCR7-CD45RA-CCR7-CD45RA+
(16). Furthermore, a recent study showed that CCR7 is
expressed on
CD27+CD45RA+CD8+
and
CD27+CD45RA-CD8+
but not on
CD27-CD45RA+CD8+
T cells (31), suggesting that CCR7 is expressed only on
naive and memory CD8+ T cells. The present study
also demonstrated that all
CD28+CD45RA+CD8+
T cells and approximately half of
CD28+CD45RA-CD8+
T cells express CCR7 while
CD28-CD45RA-CD8+
T cells and
CD28-CD45RA+CD8+
T cells do not. Analysis of CCR5 and CCR7 coexpression demonstrated
three CD28+CD45RA- memory
CD8+ T cell subsets,
CCR5+CCR7-,
CCR5-CCR7+, and
CCR5+CCR7+. Naive and
effector CD8+ T cells are
CCR5-CCR7+ and
CCR5+CCR7-,
respectively. These results indicate the following differential
lineage of these subsets in
CD28+CD45RA-CD8+
memory T cells:
CCR5-CCR7+CCR5+CCR7+CCR5+CCR7-.
From these results, we speculate the differentiation lineage of
CD8+ T cells shown in Fig. 8.
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CCR7+CD45RA-CCR7-CD45RA-CCR7-CD45RA+.
This is consistent with the results in the present study except that we
did not observe a
CCR7+CD45RA+ subset in
virus-specific CD8+ T cells. However, this does
not exclude the possibility that a
CCR7+CD45RA+ subset might
be a minor population of memory CD8+ T cells.
HCMV-specific CD8+ T cells mostly expressed CCR5,
although a small number expressed CCR7. These results strongly suggest
that the majority of HCMV-specific memory
CD8+ T cells have a
CCR5+CCR7- phenotype.
Because most HCMV-specific CD8+ T cells are
CD28-CD45RA-CD8+
and
CD28-CD45RA+CD8+
T cells, these findings together support the idea that HCMV-specific
CD8+ T cells are well-differentiated
CD8+ T cells (32). A previous study showed that CD8+CD4- thymocytes express a low level of CCR5 (7). This study together with the present study suggests that CCR5 expression is suppressed during differentiation of CD8+ thymocyte to naive CD8+ T cells and to resting memory (CCR5-CCR7+) CD8+ T cells in the periphery, and then induced after differentiation to CCR5+CCR7+ memory CD8+ T cells. The molecular mechanism of CCR5 expression during differentiation is unknown. Further studies will address the role of CCR5 in the early stages of Ag-specific CD8+ T cell differentiation.
Chemotaxis assays showed that
CCR5+CD8+ T cells migrate
in response to RANTES and MIP-1, suggesting that CCR5 expressed on
the surface of CD8+ T cells physiologically
functions as a receptor for -chemokines. Interestingly, although
CCR5 expression on
CD28-CD45RA+CD8+
T cells was much lower than on
CD28+CD45RA-CD8+
T cells and slightly lower than on
CD28-CD45RA-CD8+
T cells, the chemotactic activity of these -chemokines was highest
for CD28-CD45RA+ T cells.
The mechanism of the different effect of the -chemokines on the
different CD8+ T cell populations is unknown.
Several explanations for this may be proposed. One is that
CD28-CD45RA+CD8+
T cells express an unknown chemokine receptor for RANTES and MIP-1.
Another is that these cells have stronger systems to convert signals
via CCR5 to chemotaxis function than the other
CD8+ T cell populations. Further studies are
expected to clarify the mechanisms of -chemokine effect on the
different CD8+ T cell populations. The stronger
chemotactic response on effector
CD28-CD45RA+CD8+
T cells, which express high levels of perforin, may be important for
eradication of virus-infected cells and cancer cells.
A recent study of graft-vs-host (GVH) disease in mice showed that
administration of anti-CCR5 mAb blocks the disease
(33). This strongly suggests that alloreactive
CD8+ T cells associated with GVH disease express
CCR5 and that the effect of -chemokines on these cells is critical
for the onset of GVH disease. Blocking the interaction between
-chemokines and CCR5 with -chemokine analogs may be effective for
prevention of GVH diseases. Because HIV-1-specific
CD8+ T cells with memory, memory/effector, or
effector phenotype express CCR5, it is likely that CCR5 plays a role in
the migration of memory and effector CD8+ T cells
to inflammatory tissues where virus replicates. Thus, -chemokines
for CCR5 may be important factors in the pathogenesis of GVH disease
and in eradication of virus-infected cells.
In the present study we demonstrated functional expression of CCR5 on
Ag-specific memory and effector CD8+ T cells. Th1
CD4+ T cells also express CCR5 (6),
indicating that this receptor is expressed on T cells that are involved
in the Th1 immune response. -chemokines for CCR5 may attract Th1
CD4+ T cells as well as memory and effector
CD8+ T cells to sites of inflammation. Further
studies of CCR5 on CD8+ T cells will help clarify
the role of CCR5+CD8+ T
cells in immune responses.
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Acknowledgments |
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Footnotes |
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2 Address correspondence and reprint requests to Dr. Masafumi Takiguchi, Division of Viral Immunology, Center for AIDS Research, Kumamoto University, 2-2-1 Honjo, Kumamoto 860-0811, Japan. E-mail address: masafumi{at}kaiju.medic.kumamoto-u.ac.jp
3 Abbreviations used in this paper: MIP, macrophage-inflammatory protein; HCMV, human CMV; HCV, hepatitis C virus; GVH, graft-vs-host; MFI, mean fluorescence intensity.
Received for publication October 2, 2001. Accepted for publication December 31, 2001.
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References |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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-chemokines in human T lymphocytes. J. Immunol. 163:5763.This article has been cited by other articles:
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  T. K. Kim, L. S. St. John, E. D. Wieder, J. Khalili, Q. Ma, and K. V. Komanduri Human Late Memory CD8+ T Cells Have a Distinct Cytokine Signature Characterized by CC Chemokine Production without IL-2 Production J. Immunol., November 15, 2009; 183(10): 6167 - 6174. [Abstract] [Full Text] [PDF]
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 K Bratke, A Klug, P Julius, M Kuepper, M Lommatzsch, G Sparmann, W Luttmann, and J C Virchow Granzyme K: a novel mediator in acute airway inflammation Thorax, November 1, 2008; 63(11): 1006 - 1011. [Abstract] [Full Text] [PDF]
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R. M. Dunham, B. Cervasi, J. M. Brenchley, H. Albrecht, A. Weintrob, B. Sumpter, J. Engram, S. Gordon, N. R. Klatt, I. Frank, et al. CD127 and CD25 Expression Defines CD4+ T Cell Subsets That Are Differentially Depleted during HIV Infection J. Immunol., April 15, 2008; 180(8): 5582 - 5592. [Abstract] [Full Text] [PDF]
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D. Winter, J. Moser, E. Kriehuber, C. Wiesner, R. Knobler, F. Trautinger, P. Bombosi, G. Stingl, P. Petzelbauer, A. Rot, et al. Down-Modulation of CXCR3 Surface Expression and Function in CD8+ T Cells from Cutaneous T Cell Lymphoma Patients J. Immunol., September 15, 2007; 179(6): 4272 - 4282. [Abstract] [Full Text] [PDF]
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 M. Kallikourdis, K. G. Andersen, K. A. Welch, and A. G. Betz Alloantigen-enhanced accumulation of CCR5+ 'effector' regulatory T cells in the gravid uterus PNAS, January 9, 2007; 104(2): 594 - 599. [Abstract] [Full Text] [PDF]
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 E. S. Roberts, S. Huitron-Resendiz, M. A. Taffe, M. C. G. Marcondes, C. T. Flynn, C. M. Lanigan, J. A. Hammond, S. R. Head, S. J. Henriksen, and H. S. Fox Host response and dysfunction in the CNS during chronic simian immunodeficiency virus infection. J. Neurosci., April 26, 2006; 26(17): 4577 - 4585. [Abstract] [Full Text] [PDF]
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 D. Atanackovic, K. Pollok, C. Faltz, I. Boeters, R. Jung, A. Nierhaus, K.-M. Braumann, D. K. Hossfeld, and S. Hegewisch-Becker Patients with solid tumors treated with high-temperature whole body hyperthermia show a redistribution of naive/memory T-cell subtypes Am J Physiol Regulatory Integrative Comp Physiol, March 1, 2006; 290(3): R585 - R594. [Abstract] [Full Text] [PDF]
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 E. J.A. van Wanrooij, H. Happe, A. D. Hauer, P. de Vos, T. Imanishi, H. Fujiwara, T. J.C. van Berkel, and J. Kuiper HIV Entry Inhibitor TAK-779 Attenuates Atherogenesis in Low-Density Lipoprotein Receptor-Deficient Mice Arterioscler Thromb Vasc Biol, December 1, 2005; 25(12): 2642 - 2647. [Abstract] [Full Text] [PDF]
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 O. Gasser, A. Missiou, C. Eken, and C. Hess Human CD8+ T cells store CXCR1 in a distinct intracellular compartment and up-regulate it rapidly to the cell surface upon activation Blood, December 1, 2005; 106(12): 3718 - 3724. [Abstract] [Full Text] [PDF]
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Z. Orinska, E. Bulanova, V. Budagian, M. Metz, M. Maurer, and S. Bulfone-Paus TLR3-induced activation of mast cells modulates CD8+ T-cell recruitment Blood, August 1, 2005; 106(3): 978 - 987. [Abstract] [Full Text] [PDF]
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 T. L. Humphreys, L. A. Baldridge, S. D. Billings, J. J. Campbell, and S. M. Spinola Trafficking Pathways and Characterization of CD4 and CD8 Cells Recruited to the Skin of Humans Experimentally Infected with Haemophilus ducreyi Infect. Immun., July 1, 2005; 73(7): 3896 - 3902. [Abstract] [Full Text] [PDF]
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V. Monceaux, L. Viollet, F. Petit, R. H. T. Fang, M.-C. Cumont, J. Zaunders, B. Hurtrel, and J. Estaquier CD8+ T Cell Dynamics during Primary Simian Immunodeficiency Virus Infection in Macaques: Relationship of Effector Cell Differentiation with the Extent of Viral Replication J. Immunol., June 1, 2005; 174(11): 6898 - 6908. [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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 M. A. Kolber Impact of Immune Plasticity on Development of Cellular Memory Responses to Human Immunodeficiency Virus Type 1 Clin. Vaccine Immunol., November 1, 2004; 11(6): 1002 - 1007. [Full Text] [PDF]
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E. Lavergne, C. Combadiere, M. Iga, A. Boissonnas, O. Bonduelle, M. Maho, P. Debre, and B. Combadiere Intratumoral CC Chemokine Ligand 5 Overexpression Delays Tumor Growth and Increases Tumor Cell Infiltration J. Immunol., September 15, 2004; 173(6): 3755 - 3762. [Abstract] [Full Text] [PDF]
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 D. M. Brainard, W. G. Tharp, E. Granado, N. Miller, A. K. Trocha, X.-H. Ren, B. Conrad, E. F. Terwilliger, R. Wyatt, B. D. Walker, et al. Migration of Antigen-Specific T Cells Away from CXCR4-Binding Human Immunodeficiency Virus Type 1 gp120 J. Virol., May 15, 2004; 78(10): 5184 - 5193. [Abstract] [Full Text] [PDF]
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 D. Atanackovic, A. Block, A. de Weerth, C. Faltz, D. K. Hossfeld, and S. Hegewisch-Becker Characterization of Effusion-Infiltrating T Cells: Benign versus Malignant Effusions Clin. Cancer Res., April 15, 2004; 10(8): 2600 - 2608. [Abstract] [Full Text] [PDF]
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J. J. Zaunders, W. B. Dyer, B. Wang, M. L. Munier, M. Miranda-Saksena, R. Newton, J. Moore, C. R. Mackay, D. A. Cooper, N. K. Saksena, et al. Identification of circulating antigen-specific CD4+ T lymphocytes with a CCR5+, cytotoxic phenotype in an HIV-1 long-term nonprogressor and in CMV infection Blood, March 15, 2004; 103(6): 2238 - 2247. [Abstract] [Full Text] [PDF]
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M. X. Zhong, W. A. Kuziel, E. G. Pamer, and N. V. Serbina Chemokine Receptor 5 Is Dispensable for Innate and Adaptive Immune Responses to Listeria monocytogenes Infection Infect. Immun., February 1, 2004; 72(2): 1057 - 1064. [Abstract] [Full Text] [PDF]
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 D. Giunti, G. Borsellino, R. Benelli, M. Marchese, E. Capello, M. T. Valle, E. Pedemonte, D. Noonan, A. Albini, G. Bernardi, et al. Phenotypic and functional analysis of T cells homing into the CSF of subjects with inflammatory diseases of the CNS J. Leukoc. Biol., May 1, 2003; 73(5): 584 - 590. [Abstract] [Full Text] [PDF]
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H. Tomiyama, T. Matsuda, and M. Takiguchi Differentiation of Human CD8+ T Cells from a Memory to Memory/Effector Phenotype J. Immunol., June 1, 2002; 168(11): 5538 - 5550. [Abstract] [Full Text] [PDF]
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