Brain, Vol. 123, No. 9, 1874-1882,
September 2000
© 2000 Oxford University Press
Aberrant T cell migration toward RANTES and MIP-1
in patients with multiple sclerosis
Overexpression of chemokine receptor CCR5
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1 Multiple Sclerosis Research Laboratory, Department of Neurology and Baylor-Methodist Multiple Sclerosis Center and 2 Department of Immunology, Baylor College of Medicine, Houston, Texas, USA
Correspondence to:
Dr Jingwu Zhang, Department of Neurology, Baylor College of Medicine, 6501 Fannin Street, NB302, Houston, TX 77030, USA E-mail: jzang{at}bcm.tmc.edu
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Abstract |
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Trafficking of inflammatory T cells into the central nervous system (CNS) plays an important role in the pathogenesis of multiple sclerosis. The directional migratory ability of peripheral T cells is associated with interactions of chemokines with their receptors expressed on T cells. In this study, transmigration of peripheral T cells toward a panel of chemokines was examined in patients with multiple sclerosis and healthy individuals using Boyden chemotactic transwells. A significantly increased migratory rate preferentially toward RANTES and MIP-1
, but not other chemokines, was found in T cells obtained from multiple sclerosis patients as opposed to healthy individuals (P < 0.001). The migratory T-cell populations represented predominantly Th1/Th0 cells while non-migratory T cells were enriched for Th2-like cells. The study demonstrated further that aberrant migration of multiple sclerosis-derived T cells toward RANTES and MIP-1 resulted from overexpression of their receptors (CCR5) and could be blocked by anti-CCR5 antibodies. These findings have important implications for our understanding of the mechanism underlying aberrant T cell trafficking in multiple sclerosis. chemokines; multiple sclerosis; T cell migration
CCR5 = Type 5 CC chemokine receptor; MIP-1 = macrophage-inflammatory protein-1; MMP = matrix metalloproteinase; PBMC = peripheral blood mononuclear cells; PCR = polymerase chain reaction; PBS = phosphate-buffered saline; RANTES = normal T-cell expressed and secreted
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Introduction |
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Multiple sclerosis is a chronic inflammatory and demyelinating disease of the CNS. One of the signature histopathological features of multiple sclerosis is the focal infiltration of inflammatory cells, predominantly CD4+ Th1 cells, to the lesions (Hartung, 1993
). There is increasing evidence to suggest the CNS inflammation is associated with aberrant trafficking of pro-inflammatory T cells across the blood–brain barrier, which is composed of tight tissue junctions to prevent peripheral immune cells and large molecules from entering the CNS (Hafler et al., 1987; Chou et al., 1992; Zhang et al., 1994). Dubois and colleagues reported recently that immunization with myelin antigens failed to induce EAE (experimental autoimmune encephalo- myelitis) in young knockout mice genetically deficient in matrix metalloproteinase (MMP)-9 (Dubois et al., 1999), suggesting that access/trafficking of autoreactive T cells into the CNS is a crucial requirement for the development of EAE, an animal model for multiple sclerosis.
The ability of peripheral T cells to migrate is related to the expression of a number of T-cell surface molecules, including chemokine receptors and adhesion molecules. Many secretary molecules produced by T cells, including cytokines (e.g. 
-interferon) and MMP, also contribute to the migratory ability of T cells (Gijbels et al., 1993; Hartung et al., 1995; Hintzen et al., 1995; Hvas et al., 1997; Simpson et al., 1998). In particular, new studies have begun to unfold evidence that chemottractants (chemokines) play a critical role in aberrant T cell trafficking in patients with multiple sclerosis (Balashov et al., 1999; Sorensen et al., 1999; Strunk et al., 2000). To date, there are >40 chemokines and 10 chemokine receptors that have been identified. Expression of some chemokine receptors, such as CCR5 (receptors for RANTES, MIP-1 and MIP-1ß) and CXCR3 (receptors for IP-10 and MIG), is associated with Th1 pro-inflammatory cells, whereas Th2 anti-inflammatory cells preferentially express CCR3 (receptors for MCP-3, MCP-4 and RANTES), CCR4 (receptors for TARC and MDC) and CCR8 (Bonecchi et al., 1998; Sallusto et al., 1998; Zingoni et al., 1998). CCR5 also serves as co-receptor for HIV-1 (Deng et al., 1996; Dragic et al., 1996). It was recently reported that some chemokine receptors, including CCR5 and CXCR3, were overexpressed among lesion-derived T cells and peripheral T cells in patients with multiple sclerosis (Balashov et al., 1999; Sorensen et al., 1999; Strunk et al., 2000; reviewed in Zhang et al., 2000). Elevated levels of chemokines, IP-10, RANTES and MIG were found in CSF obtained from patients with multiple sclerosis (Sorensen et al., 1999). The studies were largely based on the binding assays in which antibodies were used to detect chemokines and chemokine receptors. These findings suggest that in multiple sclerosis, influx of pro-inflammatory T cells into the CNS may be associated with overexpression of chemokine receptors, which leads peripheral T cells to acquire aberrant trafficking properties toward certain chemokines produced at the site of pathology. However, it remains unclear as to which chemokine(s) or chemokine receptors are primarily responsible for aberrant trafficking of T cells in multiple sclerosis. This information is important for current research efforts to design specific antagonists for chemokine receptors, aiming to halt the influx of peripheral pro-inflammatory T cells into the CNS as a potential treatment for multiple sclerosis.
This study was undertaken to identify the chemokine(s) associated with aberrant T cell trafficking in multiple sclerosis using functional assays. We examined the migratory rate of peripheral T cells obtained from patients with multiple sclerosis and healthy individuals toward seven chemokines, including RANTES, MIP-1, MIG, IP-10, IL-8, MCP-1 and MCP-3. The selection of these chemokines was based on potential association of some of them and their receptors with multiple sclerosis, as reported previously based on binding studies (Balashov et al., 1999; Sorensen et al., 1999). T cell migration study was performed using standard Boyden chemotactic chambers that allow directional migration of T cells toward chemokines of interest. We describe for the first time that peripheral T cells obtained from multiple sclerosis patients as opposed to healthy individuals exhibit a unique migratory pattern preferentially toward RANTES and MIP-1 but not towards other chemokines tested. The findings correlate with overexpression of CCR5, a chemokine receptor for both RANTES and MIP-1, on peripheral T cells derived from multiple sclerosis patients compared with control T cells obtained from healthy individuals. The study has important implications for our understanding of the role of identified chemokines and chemokine receptors in aberrant trafficking of peripheral T cells in multiple sclerosis and the development of chemokine receptor antagonists as a potential treatment for multiple sclerosis.
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Material and methods |
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Reagents
All chemokines and monoclonal antibodies to CCR1, CXCR3, CCR3 and CCR5 used in this study were purchased from PharMingen (San Diego, Calif., USA). Monoclonal antibodies to CD4 and CD8 were obtained from Becton Dickinson (San Jose, Calif., USA). Medium used for cell culture and migration experiments was RPMI-1640 supplemented with 10% heat-inactivated foetal calf serum (FCS) and L-glutamine, sodium pyruvate, non-essential amino-acids and 10 mM HEPES buffer (Hyclone, Logan, UT, USA).
Human subjects
Fifteen patients with multiple sclerosis (six males and nine females) were enrolled in this study. The diagnosis was made based on clinical manifestations and laboratory studies, and confirmed by MRI (Kurtzke, 1983; Poser et al., 1983). All patients were characterized as having relapsing–remitting multiple sclerosis or chronic progressive multiple sclerosis for >2 years (average 6.2 years). The patients had not taken any immunosuppressive drugs for at least 3 months prior to the study. A group of 15 healthy volunteers (seven males and eight females of closely matched age) were included as control subjects. The protocol was approved by the Institutional Human Subjects Committee at Baylor College of Medicine. All subjects gave their informed consent to give blood samples for the study.
Enrichment of peripheral T cells
Peripheral blood mononuclear cells (PBMCs) were separated from freshly obtained blood specimens (within 2 h of blood draw) by a conventional Ficoll–Hypaque method (Zhang et al., 1992). To enrich T cells, PBMC preparations were first incubated in Petri dishes at a concentration of 2 x 106 cells/ml for 2 h at 37°C to remove monocytes. Non-adhesive cells (enriched for T cells and some B cells) were subsequently passed through a nylon wool column to enrich the T cell population (loading volume of 2 ml, cell capacity 1.5 x 108) (Hathcock et al., 1998). The nylon wool column was prepared by activating nylon wool fibres (Polysciences, Warrington, Penn., USA) with 1% HCl for 20 min and subsequently washing with water to obtain a neural pH. The column was equilibrated with RPMI-1640 and pre-warmed at 37°C for 30 min prior to use. T cells were then eluted with 15–20 ml RPMI-1640 medium. The purity of T cells was >90% as determined by flow cytometry using an anti-CD3 monoclonal antibody (Becton Dickinson).
T cell migration study
T cell migration was performed in micro-transwells of a 5 µm pore size membrane (48-well Boyden chamber, Neuro Probe, Cabin John, Md., USA) (Pilaro et al., 1990; Taub et al., 1995). T cell preparations were added at 50 000 cells/well in the upper chambers and were incubated at 37°C for 3 h. The experimental conditions were pre-determined in a series of pilot experiments. The migratory T cells in the lower chambers were numerated microscopically in three different fields. The data are expressed as the mean number of migratory T cells as a percentage of the total number of cells taken ± standard deviation.
Separation of migratory and non-migratory T cells by Boyden transwells
The same experimental conditions as described above were used for T cell migration. Briefly, T cells derived from multiple sclerosis patients and control subjects were allowed to migrate in the presence of 1 ng/ml of RANTES using large transwells of a 5 µm pore size membrane (10-well Boyden chamber, Neuro Probe). These transwells provided a large surface area to accommodate more T cells for migration. All experiments were performed under sterile conditions as the resulting T cells would be recovered and cultured for cytokine analysis. After incubation for 3 h, migratory and non-migratory T cells were collected from the lower and upper chambers, respectively. The resulting T cells were washed and plated out at 20 000 cells/well in the presence of phytohaemagglutinin (PHA) (1 µg/ml) to induce cytokine production (Kozovska et al., 1999). Culture supernatants were collected 48 h later and were subject to cytokine analysis by enzyme-linked immunosorbent assay (ELISA).
Semi-quantitative measurement of mRNA by polymerase chain reaction (PCR)
Total RNA was prepared from purified T cells using RNeasy kit (Qiagen, Valencia, Calif., USA), and cDNA was synthesized from 1 µg of isolated RNA using random hexamer and Superscript reverse transcriptase (Gibco, Grand Island, NY, USA), according to the manufacturer's instructions. PCR was performed using paired 5' and 3' primers specific for CCR5 or ß-actin. The conditions used for PCR were as follows: denaturing at 94°C for 5 min; thermal cycling at 94°C for 30 s, 57°C for 30 s, 72°C for 45 s; and an additional extension at 72°C for 7 min after the last thermal cycle. ß-actin was employed as an internal control for PCR amplification. The primer sequences are as follows: CCR5 sense primer 5'-TATTATACATCGGAGCCC, antisense primer 5'-GGTGTAATGAAGACCTTC; ß-actin sense primer 5'-AAGTACTCCGTGTGGATCGG, antisense primer 5'-AAAGCCATGCC-AAACTCATC. Thirty PCR cycles were performed to ensure that PCR amplification did not reached the plateau. The amplified PCR products were separated on 1% agarose gel and transferred to a positively charged nylon membrane (Boehringer Mannheim, Indianapolis, Ind., USA) using vacuum blot (Bio-Rad, Hercules, Calif., USA) at 5 mmHg for 90 min. DNA was fixed onto the membrane by 3 min exposure to UV crosslinking and prehybridized at 68°C for at least 1 h. Poly(A) (0.1 mg/ml) was added to prehybridization solution [5x standard saline citrate (SSC), 1% blocking solution, 0.1% N-lauroylsarcosine, 0.02% sodium dodecyl sulphate (SDS)] to reduce non-specific binding of the probe to non-target DNA. Hybridization temperature and washing conditions were optimized according to the probes to ensure a stringent hybridization condition. Hybridization was carried out in a buffer containing 5x SSC, 1% blocking solution, 0.1% N-lauroylsarcosine, 0.02% SDS, 0.3 pmol/ml digoxigenin-labelled probe for 6 h. The detection of DNA hybrid products was performed using the Digoxigenin Luminescent Detection Kit according to the manufacturer's instructions (Boehringer Mannheim). The membrane was then exposed to X-ray film for 15–30 min at room temperature. The intensities of PCR products were quantified using a Gel Doc 1000 scanning densitometer (Bio-Rad). The mRNA expression of CCR5 was analysed relative to that of ß-actin in each sample, and expressed as the ratio of CCR5 to ß-actin in percentile (Hong et al., 1999).
Flow cytometry
PBMCs were stained with the following monoclonal antibodies in pairs for direct dual staining in flow cytometric analysis: phycoerythrin (PE)-conjugated anti-CD4 and anti-CD8 antibodies/fluorescein isothiocyanate (FITC)-conjugated antibodies to CCR1, CXCR3, CCR3 and CCR5. FITC-anti-IgG1/PE-anti-IgG2a (simulset control) to detect background staining (Becton Dickinson). PBMCs were washed twice in Eppendorf tubes by the addition of 1 ml FACS buffer [phosphate-buffered saline (PBS) containing 5% foetal calf serum (FCS) and 0.01% sodium azide] and centrifugation at 2300 r.p.m. for 2 min at 4°C. The cells were then resuspended and stained with 10% of respective conjugated antibodies for 30 min on ice. After two washes the cells were resuspended in 300 µl FACS buffer and analysed by flow cytometry using a FACScan (Becton Dickinson) with gates set to read the total lymphocyte population.
Statistical analysis
A Student's t-test was used to analyse the statistical significance of the differences in the migratory rate and the expression of CCRs between the study groups. A P value of <0.05 was considered significant.
Cytokine measurement
The cytokine production of the migratory and non-migratory T cells separated using Boyden transwells was examined in culture supernatants after they were challenged with phytohaemagglutinin-P (PHA-P) (Kozovska et al., 1999). Supernatants were collected for the measurement of interferon- (IFN-), tumour necrosis factor- (TNF-), interleukin-4 (IL-4) and IL-10, 48 h after stimulation. Culture supernatants were diluted 1 : 4 with PBS prior to assays. Cytokines were determined quantitatively using ELISA kits obtained from PharMingen. The kits were used according to the manufacturer's instructions. Briefly, 96-well microtitre plates (Nunc, Maxisorp) were coated overnight at 4°C with 2 µg/well of respective mouse capturing monoclonal antibodies in PBS. Wells were then blocked at 37°C for 2 h with 2% bovine serum albumin (BSA)–PBS and washed three times with cold washing solution, containing 0.02% Tween-20. Fifty microlitres of each sample and its control were added to the adjacent wells and incubated for 2 h at ambient temperature with 50 µl of a biotinylated detecting antibody (0.25 µg/ml of each monoclonal antibody) in 2% BSA–PBS–Tween-20. Plates were washed and incubated for 30 min with streptavidin-conjugated horseradish peroxidase. One hundred microlitres of 0.0125% TMB (tetramethylbenzidine) and 0.008% H2O2 in citrate buffer was used as substrate, and colour development was stopped using 100 µl of 1 N HCl. The concentration of each cytokine in a given sample was calculated using a double standard curve of corresponding recombinant cytokine (PharMingen) in each ELISA plate, which also served as a quality control. The detection limit for all cytokine measurements was <35 pg/ml in all assays.
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Results |
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Directional migration of peripheral T cells obtained from multiple sclerosis patients and healthy individuals toward various chemokines
A group of 15 patients with either relapsing–remitting (n = 11) or chronic progressive multiple sclerosis (n = 4) was enrolled for this study. The mean expanded disability scale score (EDSS) of the patients was 3.5 with a mean disease duration of 6.2 years. Fifteen healthy volunteers were included as control subjects. Peripheral T cells purified from PBMCs were examined for random spontaneous migration (in the absence of chemokines) and directional migration toward selected chemokines using standard Boyden transwells. As shown in Fig. 1
, T cells derived from multiple sclerosis patients exhibited a significantly increased rate of random migration (16.2%) in the absence of chemokines compared with that obtained from control subjects (11.2%). When tested with a panel of seven chemokines, including RANTES, MIP-1, MIG, IP-10, IL-8, MCP-1 and MCP-3, multiple sclerosis-derived T cells displayed a characteristic migration pattern that differed from that of control T cells derived from healthy individuals (Fig. 1).
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Multiple sclerosis-derived T cells exhibited a significantly increased migratory rate preferentially toward RANTES and MIP-1
(P < 0.001) but not other chemokines. The pattern of preferential T cell migration toward RANTES and MIP-1 remained largely unchanged when the chemokines were used at 0.1 and 1 ng/ml (Fig. 1A and B
), respectively. These concentrations were found to be optimal concentrations in pilot experiments and within the concentration range of the chemokines found previously in serum and cerebrospinal fluid specimens obtained from patients with multiple sclerosis (Balashov et al., 1999; Sorensen et al., 1999). In contrast, when used at similar concentrations, the same chemokines, including RANTES and MIP-1, did not significantly alter the migratory rate of control T cells derived from healthy individuals (Fig. 1A and B). Peripheral T cells derived from selected multiple sclerosis patients and control subjects were further characterized for migration toward RANTES and MIP-1 in a dose-dependent manner. As shown in Fig. 2, the most effective concentrations of RANTES and MIP-1 were between 0.1 and 1 ng/ml for multiple sclerosis-derived T cells, while control T cells exhibited minimal migration toward the chemokines used at the same concentrations. Taken together, the findings indicate that RANTES and MIP-1 but not other chemokines, selectively alter the migratory property of multiple sclerosis-derived T cells.
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Cytokine profile of migratory T cells induced by RANTES
It was of interest to delineate whether RANTES selectively facilitated migration of a subset(s) of multiple sclerosis-derived T cells and whether the migratory T cells exhibited a characteristic cytokine profile. To address these issues, peripheral T cells were separated into migratory and non-migratory T cell populations by transwell migration toward RANTES. As illustrated in Fig. 3
, in the three multiple sclerosis-derived T cell specimens analysed, the migratory T cells derived from multiple sclerosis patients were predominantly Th1/Th0 cells while non-migratory T-cell populations were enriched for Th2-like cells. The non-migratory T cell populations produced predominantly IL-10 and substantially less -interferon compared with T cells prior to migration, suggesting that RANTES selectively facilitated transwell migration of T cells of predominantly Th1/Th0 phenotype.
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Overexpression of CCR5 in peripheral T cells obtained from multiple sclerosis patients
The observation that RANTES and MIP-1
, the ligands for CCR5, selectively altered the migratory property of multiple sclerosis-derived peripheral T cells prompted us to investigate further whether the selective effect of the two chemokines on T cell migration was related to potential overexpression of their receptors (CCR5) on multiple sclerosis-derived T cells. To this end, we first examined whether transmigration of multiple sclerosis-derived T cells induced by RANTES and MIP-1 could be blocked by an anti-CCR5 antibody. As shown in Fig. 4, the addition of an anti-CCR5 antibody, but not an isotype-matched control antibody, substantially inhibited transwell migration of multiple sclerosis-derived T cells toward RANTES and MIP-1 in a dose-dependent manner. As both RANTES and MIP-1 are known to interact with additional chemokine receptors other than CCR5, e.g. CCR3 and CCR1, it is not surprising that the blocking of CCR5 resulted in partial but not complete inhibition of T cell migration.
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Next, we examined mRNA expression of CCR5 in peripheral T cells obtained from multiple sclerosis patients and control subjects by semi-quantitative PCR. The results revealed that mRNA levels were significantly higher in multiple sclerosis-derived T cells (37.8%) that were not stimulated, compared with control T cells (15.4%, P < 0.01) (Fig. 5
). Furthermore, as illustrated in Fig. 6, when analysed for the surface expression of CCR5, multiple sclerosis-derived T cells overall exhibited a significantly higher expression of CCR5 in both CD4+ and CD8+ T-cell populations than that found in control T cells obtained from healthy individuals (P < 0.001). CD4+ T-cell preparations derived from multiple sclerosis patients were significantly enriched for CCR3+ T cells compared with control T cells (Fig. 6). In a series of additional experiments, we examined the expression of CCR1 and CXCR3 in T cells derived from 10 multiple sclerosis patients and eight healthy individuals under the same experimental conditions. The expression level of CCR1 did not differ significantly between multiple sclerosis patients and control subjects (mean 2.6 versus 2.4%, P > 0.1) while CXCR3 expression was only slightly higher in multiple sclerosis-derived T cells (16.6%) compared with control T cells (14.8%, P > 0.1) (data not shown). Taken together, these findings suggest that aberrant migration of multiple sclerosis-derived T cells toward RANTES and MIP-1 is associated with significant overexpression of their receptors (CCR5).
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Discussion |
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The recent discovery of chemokines and chemokine receptors has begun to unfold the molecular basis of how T cells are directed to the site of inflammation. The role of chemokines and chemokine receptors is particularly important in multiple sclerosis, in which myelin-destructive inflammation occurs behind the blood–brain barrier and is related to influx of peripheral pro-inflammatory T cells into the CNS. In this respect, the study described here provides direct evidence, based on functional assays, that two chemokines, namely RANTES and MIP-1
which share the same receptor (CCR5), are preferentially involved in directional migration of peripheral T cells in patients with multiple sclerosis. A concentration of 0.1 ng/ml of RANTES and MIP-1 is sufficient to induce T cell migration and mRNA synthesis of MMP-9. This concentration of RANTES and MIP-1 was found previously in CSF obtained from multiple sclerosis patients but not in control subjects (Balashov et al., 1999; Sorensen et al., 1999). Furthermore, the observation that peripheral T cells derived from multiple sclerosis patients exhibited a significantly higher random migratory rate than those from healthy individuals is consistent with the previous findings that multiple sclerosis peripheral T cells, including myelin-reactive T cells, undergo in vivo activation and have migratory advantages (Hafler et al., 1987; Allegretta et al., 1990; Chou et al., 1992; Zhang et al., 1994). The study described here provides experimental evidence that RANTES selectively facilitated migration of Th1/Th0-like cells. The finding is particularly relevant to the understanding of the mechanism underlying trafficking of pro-inflammatory T cells into the CNS and the site of pathology as the pro-inflammatory cytokines produced by Th1 cells, such as TNF- and -interferon, are associated with clinical activity in multiple sclerosis (Panitch et al., 1987; Sharief et al., 1991).
It was demonstrated in this study that aberrant migration of multiple sclerosis-derived T cells preferentially toward RANTES and MIP-1 is attributable to the overexpression of CCR5. Overexpression of CCR5 in PBMCs obtained from patients with multiple sclerosis was also reported by other investigators based on binding assays (Balashov et al., 1999; Sorensen et al., 1999; Strunk et al., 2000). It is important to note that although several other chemokines (e.g. IP-10) and chemokine receptor expression (e.g. CXCR3) are also elevated in the CSF, peripheral blood and even in post-mortem multiple sclerosis lesions as reported previously, they do not directly alter the migratory property of multiple sclerosis-derived T cells in the functional studies described here. It suggests that local inflammatory processes may result in the production and accumulation of a variety of chemokines, but not all chemokines produced at the site of inflammation are necessarily associated with aberrant T cell trafficking in multiple sclerosis, strengthening the importance of the finding described here based on the functional assays. On the other hand, it is conceivable that CCR5 may not be the only chemokine receptor involved in the aberrant T cell migration in multiple sclerosis. This possibility is supported by a previous report that the lack of CCR5 expression fails to protect against multiple sclerosis (Bennetts et al., 1997) and by the observation that blocking of CCR5 did not result in complete inhibition of T cell migration, as described here. As some of the chemokines, including RANTES and MIP-1, share different receptors, blocking of a single chemokine receptor is not sufficient to abolish T cell migration.
The study suggests that overexpression of CCR5 is critical to aberrant migration of peripheral T cells toward the site of inflammation in multiple sclerosis. It is probable that the observed overexpression of CCR5 in multiple sclerosis patients may be triggered by certain events associated with multiple sclerosis. Among many possibilities, viral infection is of particular relevance. There is increasing evidence to suggest that human herpes virus (HHV)-6 may be associated with multiple sclerosis. Cell-free viral DNA and elevated antibody titers to HHV-6 were found in CSF and serum specimens of patients with multiple sclerosis but not in healthy individuals (Soldan et al., 1997). It was reported recently that an open-reading frame within HHV-6, designated as U12 gene, encodes a functional receptor for RANTES (Isegawa et al., 1998). Although it is unclear at this time whether re-activation of HHV-6 in multiple sclerosis patients is responsible for the overexpression of CCR5 per se or the induction of a viral chemokine receptor similar to CCR5, the finding opens new possibilities in the search for a link between viral infection and the autoimmune mechanism involved in multiple sclerosis.
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Notes |
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* These authors contributed equally to the work
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Acknowledgments |
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This work was supported in part by Biogen Inc. and the Albert & Ethel Herzstein Foundation.
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References |
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TopAbstractIntroductionMaterial and methodsResultsDiscussionReferences |
|---|
Allegretta M, Nicklas JA, Sriram S, Albertini RJ. T cells responsive to myelin basic protein in patients with multiple sclerosis. Science 1990; 247: 718–21.
Balashov KE, Rottman JB, Weiner HL, Hancock WW. CCR5(+) and CXCR3(+) T cells are increased in multiple sclerosis and their ligands MIP-1alpha and IP-10 are expressed in demyelinating brain lesions. Proc Natl Acad Sci USA 1999; 96: 6873–8.
Bennetts BH, Teutsch SM, Buhler MM, Heard RN, Stewart GJ. The CCR5 deletion mutation fails to protect against multiple sclerosis. Hum Immunol 1997; 58: 52–9.[ISI][Medline]
Bonecchi R, Bianchi G, Bordignon PP, D'Ambrosio D, Lang R, Borsatti A, et al. Differential expression of chemokine receptors and chemotactic responsiveness of type 1 T helper cells (Th1s) and Th2s. J Exp Med 1998; 187: 129–34.
Chou YK, Bourdette DN, Offner H, Whitham R, Wang RY, Hashim GA, et al. Frequency of T cells specific for myelin basic protein and myelin proteolipid protein in blood and cerebrospinal fluid in multiple sclerosis. J Neuroimmunol 1992; 38: 105–13.[ISI][Medline]
Deng H, Liu R, Ellmeier W, Choe S, Unutmaz D, Burkhart M, et al. Identification of a major co-receptor for primary isolates of HIV-1. Nature 1996; 381: 661–6.[Medline]
Dragic T, Litwin V, Allaway GP, Martin SR, Huang Y, Nagashima KA, et al. HIV-1 entry into CD4+ cells is mediated by the chemokine receptor CC-CKR-5. Nature 1996; 381: 667–73.[Medline]
Dubois B, Masure S, Hurtenbach U, Paemen L, Heremans H, van den Oord J, et al. Resistance of young gelatinase B-deficient mice to experimental autoimmune encephalomyelitis and necrotizing tail lesions. J Clin Invest 1999; 104: 1507–15.[ISI][Medline]
Gijbels K, Proost P, Masure S, Carton H, Billiau A, Opdenakker G. Gelatinase B is present in the cerebrospinal fluid during experimental autoimmune encephalomyelitis and cleaves myelin basic protein. J Neurosci Res 1993; 36: 432–40.[ISI][Medline]
Hafler DA, Weiner HL. In vivo labeling of blood T cells: Rapid traffic into cerebrospinal fluid in multiple sclerosis. Ann Neurol 1987; 22: 89–93.[ISI][Medline]
Hartung HP. Immune-mediated demyelination. Ann Neurol 1993; 33: 563–7.[ISI][Medline]
Hartung HP, Reiners K, Archelos JJ, Michels M, Seeldrayers P, Heidenreich F, et al. Circulating adhesion molecules and tumor necrosis factor receptor in multiple sclerosis: correlation with magnetic resonance imaging. Ann Neurol 1995; 38: 186–93.[ISI][Medline]
Hathcock K. T cell enrichment by non-adherence. In: Coligan JE, Kruisbeek AM, Margulies DH, Shevach EM, Strober W, editors. Current protocols in immunology, Vol. 1. New York: John Wiley; 1998; p. 321–4.
Hintzen RQ, Fiszer U, Fredrikson S, Rep M, Polman CH, van Lier RA, et al. Analysis of CD27 surface expression on T cell subsets in MS patients and control individuals. J Neuroimmunol 1995; 56: 99–105.[ISI][Medline]
Hong J, ZangYC, Tejada-Simon MV, Kozovaska M, Li S, Singh R, et al. A common TCR V-D-J sequence in V beta 13.1 T cells recognizing an immunodominant peptide of myelin basic protein in multiple sclerosis. J Immunol 1999; 163: 3530–8.
Hvas J, McLean C, Justesen J, Kannourakis G, Steinman L, Oksenberg JR, et al. Perivascular T cells express the pro-inflammatory chemokine RANTES mRNA in multiple sclerosis lesions. Scand J Immunol 1997; 46: 195–203.[ISI][Medline]
Isegawa Y, Ping Z, Nakano K, Sugimoto N, Yamanishi K. Human herpesvirus 6 open reading frame U12 encodes a funcitional beta-chemokine receptor. J Virol 1998; 72: 6104–12.
Kozovska ME, Hong J, Zang YC, Li S, Rivera VM, Killian JM, et al. Interferon beta induces T-helper 2 immune deviation in MS. Neurology 1999; 53: 1692–7.
Kurtzke JF. Rating neurologic impairment in multiple sclerosis: an Expanded Disability Status Scale (EDSS). Neurology 1983; 33: 1444–52.
Panitch HS, Hirsch RL, Schindler J, Johnson KP. Treatment of multiple sclerosis with gamma interferon: exacerbations associated with activation of the immune system. Neurology 1987; 37: 1097–102.
Pilaro A, Sayers TJ, McCormick KL, Reynold CW, Wiltrout RH. An improved in vitro assay to quantitate chemotaxis of rat peripheral blood large granular lymphocytes (LGL). J Immunol Methods 1990; 135: 213–23.[ISI][Medline]
Poser CM, Paty DW, Scheinberg L, McDonald WI, Davis FA, Ebers GC, et al. New diagnostic criteria for multiple sclerosis: guidelines for research protocols. Ann Neurol 1983; 13: 227–31.[ISI][Medline]
Sallusto F, Lenig D, Mackay CR, Lanzavecchia A. Flexible programs of chemokine receptor expression on human polarized T helper 1 and 2 lymphocytes. J Exp Med 1998; 187: 875–83.
Sharief MK, Hentges R. Association between tumor necrosis factor-alpha and disease progression in patients with multiple sclerosis. N Engl J Med 1991; 325: 467–72.[Abstract]
Simpson JE, Newcombe J, Cuzner ML, Woodroofe MN. Expression of monocyte chemoattractant protein-1 and other beta-chemokines by resident glia and inflammatory cells in multiple sclerosis lesions. J Neuroimmunol 1998; 84: 238–49.[ISI][Medline]
Soldan SS, Berti R, Salem N, Secchiero P, Flamand L, Calabresi PA, et al. Association of human herpes virus 6 (HHV-6) and multiple sclerosis: increased IgM response to HHV-6 early antigen and detection of serum HHV-6 DNA. Nat Med 1997; 3: 1394–7.[ISI][Medline]
Sorensen TL, Tani M, Jensen J, Pierce V, Lucchinetti C, Folcik VA, et al. Expression of specific chemokines and chemokine receptors in the central nervous system of multiple sclerosis patients. J Clin Invest 1999; 103: 807–15.[ISI][Medline]
Strunk T, Bubel S, Mascher B, Schlenke P, Kirchner H, Wandinger KP. Increased numbers of CCR5+ interferon-gamma- and tumor necrosis factor-alpha-secreting T lymphocytes in multiple sclerosis patients. Ann Neurol 2000; 47: 269–73.[ISI][Medline]
Taub DD, Key ML, Clark D, Turcovski-Corrales SM. Chemotaxis of T lymphocytes on extracellular matrix proteins. J Immunol Methods 1995; 184: 187–98.[ISI][Medline]
Zhang GX, Baker CM, Kolson DL, Rostomi AM. Chemokines and chemokine receptors in the pathogenesis of multiple sclerosis. Mult Scler 2000, 6: 3–13.
Zhang J, Medaer R, Hashim GA, Ying C, van den Berg-Loonen E, Raus JC. Myelin basic protein-specific T lymphocytes in multiple sclerosis and controls: precursor frequency, fine specificity, and cytotoxicity. Ann Neurol 1992; 32: 330–8.[ISI][Medline]
Zhang J, Markovic-Plese S, Lacet B, Raus J, Weiner HL, Hafler DA. Increased frequency of interleukin 2-responsive T cells specific for myelin basic protein and proteolipid protein in peripheral blood and cerebrospinal fluid of patients with multiple sclerosis. J Exp Med 1994; 179: 973–84.
Zingoni A, Soto H, Hedrick JA, Stoppacciaro A, Storlazzi CT, Sinigaglia F, et al. The chemokine receptor CCR8 is preferentially expressed in Th2 but not in Th1 cells. J Immunol 1998; 161: 547–51.
Received February 16, 2000. Revised May 12, 2000. Accepted May 15, 2000.
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