The etiology of Rheumatoid Arthritis (RA) remains unclear despite the demonstration of numerous immunological aberrations in this disease. Until recently, the majority of investigations of the cellular immune component in RA have been concerned with immune function of cells derived from the peripheral blood. However, it is becoming increasingly clear that the site of pathology in RA, the synovial space (SS), shows a cellular immune response that is not just a simple reflection of the cellular immune component in the peripheral blood. Investigation of cellular immunity in the SS has been somewhat difficult owing to the relatively few numbers of cells that can be obtained from this site. In addition the heterogeneous composition of the few cells obtainable has made investigations of the specificity and function of such cells difficult. Such obtacles should now be surmountable due to the recent increase in information and technology relating to T cell cloning and long-term In Vitro growth of T cells with various growth factors. The investigations outlined in this proposal involve cloning and expanding SS T cells into large sized monoclonal populations and examining the antigen specificity and function of such SS derived T cell clones. T cells isolated from the synovium and synovial fluid (SS) of RA patients and patients with other types of arthritis will be cloned with and without a preculture with antigens of putative significance in RA. The cloning will be accomplished either on soft agar or in limiting dilution cultures. The resulting clones will be expanded with the addition of T cell growth factor (TCGF) or Interleukin 2) and eventually tested for antigen specificity using tritiated thymidine incorporation proliferation assays. SS T cell clones will also be sought that suppress EBV induced B cell proliferation. Comparisons of the antigen specific T cell repertoire in the SS versus that in the peripheral circulation will be made. The antigen specific repertoire in the SS of RA patients versus that in the SS of non-RA arthritis patients will be compared. Cloning of T cells from RA-SS will allow for a new approach to the investigation of RA and autoimmune diseases in general.
