To study the selective pressures responsible for the expansion of T cells in rheumatoid arthritis, we constructed cDNA mini-libraries from purified CD4+ and CD8+ T cells prepared from peripheral blood and from synovial fluids of two rheumatoid arthritis patients. Comparison of these libraries by hybridization with specific probes indicated that V beta 2 and V beta 8 transcripts are selectively enriched in the CD4+ synovial fluid lymphocyte population, while V beta 4 was over-represented among both the CD4+ and CD8+ subsets. The enrichment of V beta 14 and V beta 17 observed in synovial fluid T cells of one patient was, however, selectively confined to the CD8+ T-cell subpopulation. Sequence analysis of several V beta 2, V beta 4 and V beta 8 clones, derived from CD4+ cells, revealed a high degree of heterogeneity in the V beta-D beta-J beta junctions, while a more biased utilization of J segments and a more restricted junctional heterogeneity were observed in V beta 4, V beta 14 and V beta 17 clones derived from CD8+ cells. These data suggest that the disease may be induced by the initial activation of a rather heterogeneous population of T-helper cells that are later responsible for the expansion of a more restricted pool of highly specific effector lymphocytes.