Molecular characterization of the V_γ9 T cell receptor repertoire expressed in patients with rheumatoid arthritis

Abstract

We have characterized the variable (V)gamma 9 T cell receptor (TcR) repertoire expressed in synovial membrane (SMC) and peripheral blood mononuclear cells (PBMC) of five rheumatoid arthritis (RA) patients. The experimental approach was to sequence the junctional regions of polymerase chain reaction (PCR)-amplified V gamma 9 transcripts; gamma chain sequences were compared with those found in normal PBMC. For normal PBMC, a large proportion of V gamma 9 transcripts used the joining (J)gamma P gene segment whereas a few used J gamma 2. Despite this restriction in J gamma usage, there was extensive junctional region diversity with a unique sequence observed in each transcript examined. In contrast to normal PBMC, J gamma usage of V gamma 9 transcripts in PBMC of two patients was skewed towards J gamma 2. This deviation in J gamma usage was more pronounced in SMC since all patients expressed V gamma 9 transcripts in SMC which predominantly used J gamma 2, as opposed to J gamma P in normal PBMC. Further, approximately 60% of V gamma 9 transcripts in PBMC of each of three patients had identical junctional region sequences, although the specific sequences were unique in each patient. For two of these patients the dominant transcript found in the PBMC was detected in the corresponding SMC at about 10% and 40%, respectively. Overall, our findings indicate that V gamma 9-bearing T cells in RA peripheral blood are largely derived from clonal expansion whereas in the synovium there is expression of a population of these cells that are mainly polyclonal. This may reflect in vivo expansion in response to a V gamma 9/J gamma 2 region-specific antigen. The data presented in this report suggest that V gamma 9+ T cells may play a role in the development of RA.

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

“…The study had appropriate institutional ethical approval. Patients 1 , 2 , 3 , 4 and 5 in this study correspond respectively with patients 1 , 2 , 3 , 6 and 7 in our earlier study of analysis of V gene usage of y/6 TcR expressed in RA patients [13]. HLA-DR types of patients 1 to 5 are DR7,DRwlO; DR3,DR4 (Dw14); DR2,DR4 (Dw4); DR1,DR7; DR1,-, respectively.…”

“…y/6 T cells in the synovial fluid and synovial membrane of RA patients have been shown to express predominantly Val chains [ll-131 and to a lesser extent Vb2 chains [13]. This is in contrast to human PBMC which express mainly V62 chains, preferentially paired with TcR y chains encoded by the V,9 gene [14].…”

“…First-strand cDNA was synthesized, as described elsewhere [13], from total RNA isolated from SMC and PBMC by the guanidinium thiocyanate-phenol-chloroform extraction method [ 191. Approximately 5% of single-stranded cDNA template was amplified in 50 pl reactions containing 1 x PCR buffer (10 mM Tris-HC1 (pH8.4), 50 mM KC1,2.5 mM MgC12, 0.01% gelatin), 0.2 mM of each dNTP, 25 pmol 5 ' V~l (5'-TCTGGATCAAGTGTGGC-3') or Vh2 (5'-TC-TGGGCAGGAGTCATGT-3') sense primer, 25 pmol of 3' constant region gene antisense primer C62 (5'-TTCACCA-GACAAGCGACA-3') and 1.25 U Taq DNA polymerase (Promega, Madison, WI).…”

Cellular Interactions of Synovial Fluid γδ T Cells in Juvenile Idiopathic Arthritis

“…Although the precise etiology is still unknown, JIA is an autoimmune disease, and T cells are thought to be key players in this process (12). Interestingly, although the Vg9Jg1.2 gene rearrangement is less commonly expressed among synovial fluid (SF) than peripheral blood (PB) gd T cells, IPP was recently found to induce expansion of SF Vg9 + T in virtually all forms of JIA (13,14). However, despite the known proinflammatory potential of Vg9 + T cells in healthy individuals, higher levels of expansion were associated with clinical remission in these patients, suggesting that IPP-induced activation of the cells in some unknown manner contributes to control synovial inflammation (13).…”

γδ T cell receptor gene expression by muscle-infiltrating lymphocytes in the idiopathic inflammatory myopathies

“…Likewise, similar studies describing restricted patterns of 7^ TCR gene expression suggest an important role for -yS T cells in many human autoimmune diseases such as multiple sclerosis [13,16.26] and rheumatoid arthritis [12,17,27,28]. Unfortunately, many of these studies have yielded conflicting data on the extent and nature of 71^ TCR expression in inflammatory lesions [15,16,19,[28][29][30][31]. In the IIM, immunohistochemical analyses of muscle biopsies suggest that -yS T cells are an infrequent component of lymphocytic infiltrates among most patients surveyed [9,32].…”