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γδ T Cell Receptor Analysis Supports a Role for HSP 70 Selection of Lymphocytes in Multiple Sclerosis Lesions

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Abstract

Background

Interactions between γδ T cells and heat shock proteins (HSP) have been proposed as contributing factors in a number of diseases of possible autoimmune etiology but definitive evidence to support this hypothesis has been lacking. In multiple sclerosis (MS), a chronic inflammatory neurologic disease, HSP and γδ T cells are known to colocalize in brain lesions. Analysis of T cell receptor (TCR) gene usage in these lesions has detected evidence of clonality within both the Vδ2-Jδ1 and Vδ2-Jδ3 populations of γδ T cells. In our own studies, using direct sequence analysis, a dominant Vδ2-Jδ3 TCR sequence was found in 9 MS brain samples, suggesting a response to a common antigen. In this report, we have examined γδ T cell receptor gene usage in MS peripheral blood T cell lines selected for reactivity to HSP 70.

Materials and Methods

TCR rearrangement patterns for Vδ2-Jδ1 and Vδ2-Jδ3 were studied using the polymerase chain reaction (PCR) and a direct sequencing technique in populations of peripheral blood mononuclear cells (PBMC) cultured with Mycobacterium tuberculosis (M. tuberculosis) purified protein derivative (PPD) and then selected for reactivity to a 70-kD heat shock protein (HSP70). Cells were obtained from healthy donors, patients with MS, and patients with tuberculosis (TB). PCR products were subjected to direct sequence analysis to look for evidence of clonality within these T cell lines and to define the sequence of the V-D-J (CDR3) region of the TCR.

Results

In freshly isolated PBMC, both Vδ2-Jδ1 and Vδ2-Jδ3 gene rearrangement patterns were detected, whereas in HSP70+ T cell lines the predominant δ chain rearrangement pattern was Vδ2-Jδ3. Direct sequence analyses indicated that in cells reactive with HSP70 the Vδ2-Jδ3 sequences were usually oligoclonal and used Dδ3 exclusively. In four of four MS and two of three TB patients, the oligoclonal sequences in the HSP70+ T cell lines were identical to one another and to a dominant sequence previously detected in MS brain lesions. In two of three HSP70+ T cell lines from healthy controls, the oligoclonal sequences differed from those found in both groups of patients but were identical to one another except for a small region of heterogeneity in the second N region. In contrast, in freshly isolated PBMC or in PPD+HSP70− T cell lines, the Vδ2-Jδ3 gene rearrangement patterns were usually polyclonal and dominant sequences were rarely identified.

Conclusions

These results support the conclusion that a subpopulation of γδ T cells in MS lesions are responding to HSP 70 and that non-CNS-specific antigens contribute to the pathogenesis of MS.

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Acknowledgments

Supported in part by USPHS Grants NS 11920 and NS 08952, and RG 1001-H-8 and FA 1095 from the National Multiple Sclerosis Society, the New York Community Trust and the Gladstein Foundation; and by grants from the Italian Multiple Sclerosis Society (AISM) and the Istituto Mediterraneo per la Ricerca Scientifica, Pozzilli, Italy. Presented in part at the IVth International Congress of Neuroimmunology, Amsterdam, The Netherlands, 1994.

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Authors and Affiliations

  1. Department of Pathology, Albert Einstein College of Medicine, 1300 Morris Park Avenue, Bronx, New York, 10461, USA

    Luca Battistini, Cedric S. Raine & Celia F. Brosnan

  2. Department of Neuroscience, Albert Einstein College of Medicine, Bronx, New York, USA

    Cedric S. Raine & Celia F. Brosnan

  3. Department of Neurology, Albert Einstein College of Medicine, Bronx, New York, USA

    Cedric S. Raine

  4. I Clinica Neurologica, Universita “La Sapienza”, Rome, Italy

    Luca Battistini, Marco Salvetti, Giovanni Ristori & Marika Falcone

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  1. Luca Battistini
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Battistini, L., Salvetti, M., Ristori, G. et al. γδ T Cell Receptor Analysis Supports a Role for HSP 70 Selection of Lymphocytes in Multiple Sclerosis Lesions. Mol Med 1, 554–562 (1995). https://doi.org/10.1007/BF03401592

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