We propose to study the clonal heterogeneity of T lymphocytes in autoimmune diseases, particularly in rheumatoid arthritis. This will be carried out through an analysis of the T cell receptors (TCR). Framework monoclonal antibodies (mAb) against the TCR delta chain (beta F1) and against the TCR beta chain (anti-TCR delta 1) will be used to study the TCR alpha beta and TCR gamma delta proteins from rheumatoid arthritis T cells. The T cells will include the activated T cells from peripheral blood, as well as T cells from synovial fluid and synovial tissue. TCR protein will be characterized by two dimensional gel analyses. TCR gene rearrangements will be examined in Southern blotting analyses. TCR protein will then be isolated for frequently immunization of mice in the production of mAb that may recognize frequently used or cross-reactive TCR gene segment encoded determinants. Complementary used or cross-reactive TCR gene segment encoded determinants. DNA clones corresponding to these gene segments will be cloned and sequenced. Analysis on a more limited basis will be carried out in type 1 diabetes and systemic lupus erythematosus. The framework mAb against the TCR will also be evaluated as diagnostic tools for distinguishing malignant versus benign T- lineage proliferations, based on the discordant expression of CD3 and TCR proteins in neoplasia. All analyses performed will include both receptors, TCR alpha beta and TCR gamma delta. We hope that the proposed studies will help us to understand the role of T cells in autoimmune diseases and that T cell receptors may prove useful in the diagnosis of T-lineage neoplasms.
