Analysis of T cell receptor V_α and V_β gene usage in synovia of patients with rheumatoid arthritis

Abstract

Summary T cells are thought to play a fundamental role in the pathogenesis of rheumatoid arthritis (RA), Activated T cells expressing α/β T cell receptor (Tcr) infiltrate the rheumatoid synovium and could potentially initiate a local inflammatory response directed against joint constituents, A Tcr repertoire with restricted heterogeneity may reflect a selective expansion of T cells reactive to a few antigenic determinants within the synovium. To determine whether predominant Vα and/or Vβ gene usage of the expressed α/β Tcr repertoire is a feature of synovial T cells in patients with RA, the polymerase chain reaction (PCR) was used to amplify Tcr‐α and Tcr‐β chain transcripts. The peripheral blood and synovia of five patients with adult RA were examined and no evidence of preferential use of 19 Tcr Vα gene families was found. Similarly, most of the 18 Tcr Vβ gene families could be detected in RA synovia although there were quantitative differences in Tcr Vβ gene expression when compared to peripheral blood. This report shows that when the extremely sensitive assay of oligonucleotide hybridization of PCR amplified Tcr transcripts is used, permitting identification of specific V gene families, the α/β Tcr repertoire in the rheumatoid synovium is more diverse than was previously thought. Therefore, in patients with RA of long duration, the synovial T cell response is most likely to be polyclonal.

T cell contributions to the different phases of granuloma formation

“…Furthermore, injection of a GFP-tagged CNS-autoantigen-specific T cell line into BCG-infected mice results in the localization of GFP positive T cells in the granuloma [8]. The presence of non-specific T cells occurs with other types of chronic inflammation, such as that which occurs in autoimmune disease models [55][56][57].…”

Evidence for oligoclonality of T cell receptor δ chain transcripts expressed in rheumatoid arthritis patients

“…Mononuclear cells were isolated from peripheral blood (PBMC) and synovium (SMC) of patients, as described previously [18]. SMC and PBMC of patients were not cultured or stimulated in vitro.…”

Variable gene usage of T cell receptor γ- and δ-chain transcripts expressed in synovia and peripheral blood of patients with rheumatoid arthritis

“…Synovium was collected into HBSS and was mechanically minced, before enzymatic digestion. The sample was then filtered and mononuclear cells (SMC) were separated on a Ficoll gradient, as described elsewhere [18].…”

“…Synovium was collected into HBSS and was mechanically minced, before enzymatic digestion. The sample was then filtered and mononuclear cells (SMC) were separated on a Ficoll gradient, as described elsewhere [18]. Controls PBMC were isolated from heparinized blood obtained from nine healthy donors, by density gradient centrifugation using Lymphoprep (Nycomed, AS, Norway).…”