Identification of clonally expanded T cells in rheumatoid arthritis using a sequence enrichment nuclease assay.

Abstract

Identification of expanded clones engaged in immune and autoimmune responses is still imperfect, since they are often diluted by irrelevant cells expressing diverse specificities. To efficiently characterize T cell receptors expressed by clonally expanded lymphocytes in rheumatoid arthritis (RA) and other inflammatory conditions, we developed an assay system, termed sequence enrichment nuclease assay (SENA). Key elements of SENA are the efficiency of heat-denatured DNA strand reassociation, which increases exponentially with concentration, and the elimination of unhybridized sequences by single-strand-specific DNase. T cell clonal expansions were identified primarily in synovial fluids, but also in peripheral blood of RA patients. Synovial fluids had more prominent expansions in the CD8 than the CD4 subset, whereas clonal expansions in the CD4 subset predominated among peripheral blood lymphocytes.

Pathologic clonal cytotoxic T-cell responses: nonrandom nature of the T-cell–receptor restriction in large granular lymphocyte leukemia

“…Such processes may include, for example, typical aplastic anemia 53 and classic autoimmune diseases such as rheumatoid arthritis and multiple sclerosis. [54][55][56][57] Of note is that in all of these conditions, evolution of LGL leukemia has been described, 45,47,[58][59][60] and the LGL clone may have evolved from an initially polyclonal process. The TCR in LGL leukemia had been investigated at DNA sequence level in only a limited number of cases, 54,59,[61][62][63] and perhaps not surprisingly (due to diversity of the HLA background of patients) identical specificities were not found.…”

Skewed T-cell receptor BV14 and BV16 expression and shared CDR3 sequence and common sequence motifs in synovial T cells of rheumatoid arthritis

“…[3][4][5] Further supporting evidence includes skewing of cytokine environment in favor of T cell-mediated inflammation and clonal expansion of infiltrating T cells in the affected joints. [6][7][8][9][10][11][12] However, the antigen specificity of the infiltrating T cells in rheumatoid synovium is unknown. Several self-antigens, including collagen type II, heat-shock proteins and others, are implicated in RA based on T-cell reactivity to these antigens in patients with RA.…”

CD4 and CD8 T Cells in Susceptibility/Protection to Collagen-Induced Arthritis in HLA-DQ8-Transgenic Mice: Implications for Rheumatoid Arthritis

“…These include 1) the infiltration of inflamed synovium with predominantly T cells, 2) improvement in joint disease manifestations of arthritis following treatment with depleting CD4 mAb, and 3) multiple oligoclonally expanded CD4 T cells within the rheumatoid joint (11)(12)(13)(14)(15). Recent studies suggest that RA patients carry expanded CD4 clonotypes, which are not restricted to the joint but are also found in periphery, thus indicating a systemic nature of the disease (16,17). The observation that CD8 clonotypes are detected in synovial fluids while CD4 ϩ clonal expansions have been found predominantly in periphery has led to the notion that CD8 T cells may play a significant role in RA pathogenesis (17,18).…”

“…Recent studies suggest that RA patients carry expanded CD4 clonotypes, which are not restricted to the joint but are also found in periphery, thus indicating a systemic nature of the disease (16,17). The observation that CD8 clonotypes are detected in synovial fluids while CD4 ϩ clonal expansions have been found predominantly in periphery has led to the notion that CD8 T cells may play a significant role in RA pathogenesis (17,18). T cells lacking both CD4 and CD8 surface markers have also been isolated from synovial fluid of RA patients (19).…”