HLA‐B27‐Restricted Cytotoxic T Lymphocyte Responses to Arthritogenic Enterobacteria or Self‐Antigens are Dominated by Closely Related TCRBV Gene Segments. A Study in Patients with Reactive Arthritis

Abstract

Identification of the T-cell receptors (TCR) used by synovial cytotoxic T lymphocytes (CTL) of patients with reactive arthritis (ReA) may be crucial to better understanding the pathogenetic mechanism underlying the HLA-B27 association of spondylarthropathies. The authors, therefore, sequenced 25 TCRB chains from HLA-B27-restricted CD8+ CTL clones and two clonal lines specific for self- or Yersinia enterocolitica antigen isolated from synovial fluids of 3 HLA-B27+ patients with ReA and PBL of one healthy HLA-B27+ individual. Fourteen non-HLA-B27-restricted CTL served as controls. Both autoreactive and Y. enterocolitica specific HLA-B27-restricted CTL used a highly limited set of VB genes with preferential rearrangement of three closely related VB families (VB 13, 14, 17), suggesting that these families contain a preferred site for contact with the HLA-B27 molecule. In addition, the presence of limited TCRBJ usage, limited heterogeneity in CDR3 sequences and dominant clones from individual donors among these CTL indicate that TCRB chain usage is further restricted by a limited set of peptides bound to the HLA-B27 molecule. Limited TCR usage by SF CTL of ReA patients may lay a basis for therapeutical manipulation of the T-cell response in the spondylarthropathies.

Detection of clonally rearranged T-cell-receptor gamma chain genes from T-cell malignancies and acute inflammatory rheumatic disease using PCR amplification, PAGE, and automated analysis

“…In Yersinia-and Salmonella-induced reactive arthritis, HLA B27-restricted T-lymphocyte clones from synovial fluid, which may provide the missing link between the antecedent bacterial infection and the genetic association (HLA B27) of postinfectious seronegative spondylarthropathies [3], have been isolated. In Yersinia enterocolitica-induced reactive arthritis, HLA B27-restricted CTL used a highly limited set of Vb genes ( Vb 13,14,17), suggesting that these families contain a preferred site of contact with the HLA B27 molecule [3]. Also heterogeneity in CDR 3 sequences was limited, suggesting that Vb usage is further restricted by a limited set of peptides presented together with the HLA B27 molecule.…”

CD4+ and CD8+ clonal T cell expansions indicate a role of antigens inankylosing spondylitis; a study in HLA-B27+ monozygotic twins

“…Total cellular RNA was extracted from 3–5 × 10 6 cells (PBMC, purified CD4 + T cells, purified CD8 + T cells) using the RNeasy TM RNA extraction Kit (Qiagen, Hilden, Germany) according to the manufacturer's recommendations. As previously described [9], 1 μ g of RNA from each sample was converted to cDNA in a volume of 75 μ l reaction mixture containing oligo dT (Sigma, Deisenhofen, Germany), M‐MLV reverse transcriptase (G ibco BRL, Eggenstein, Germany), RNasin (Promega, Madison, WI) and dATP, dTTP, dGTP, dCTP (Boehringer, Mannheim, Germany) at 125 μ m each in 50 m m Tris–HCl pH 8·3, 75 m m KCl, 3 m m MgCl 2 , 10 m m DTT (G ibco ).…”

Pathogenesis of reactive arthritis

“…A central role for CD8+ T cells is also supported by the finding of an oligoclonal expansion among CD8+ T cells in the synovial fluid of HLA-B27-positive patients with ReA [48][49][50]. A high homology of T-cell receptors and an even identity was found in different patients with ReA triggered by different bacteria in these studies, suggesting that similar yet unidentified antigens are seen by these CD8+ T cells.…”