Detection of T Cell Receptors in Early Rheumatoid Arthritis Synovial Tissue

• Published in: Pathobiology (Basel) Vol. 63; no. 2; pp. 100 - 108
• Main Authors: Ramanujam, Thaila, Luchi, Monica, Schumacher, Ralph, Zwillich, Samuel, Chang, Chang Pei, Callegari, Peter E., Von Feldt, Joan M., Fang, Qiong, Weiner, David B., Williams, William V.
• Format: Journal Article
• Language: English
• Published: Basel, Switzerland 1995
• Subjects: Adult; Aged; Aged, 80 and over; Amino Acid Sequence; Arthritis, Rheumatoid - immunology; Base Sequence; Blotting, Southern; DNA, Complementary; Female; Humans; Male; Middle Aged; Molecular Sequence Data; Original Paper; Polymerase Chain Reaction; Receptors, Antigen, T-Cell, alpha-beta - analysis; Receptors, Antigen, T-Cell, alpha-beta - chemistry; Receptors, Antigen, T-Cell, alpha-beta - genetics; Synovial Membrane - immunology; Synovitis - immunology; Polymerase chain reaction; Early arthritis; Synovial tissue; Synovial biopsy; T cell receptor; Rheumatoid arthritis
• ISSN: 1015-2008; 1423-0291
• DOI: 10.1159/000163940

Synovial tissue is rarely available from patients with early synovitis, with the exception of synovial biopsies. However, T cell populations early in the development of synovitis may be enriched in antigen-specific cells and critical to disease pathogenesis. To investigate the T cell repertoire in early synovitis, we utilized a PCR protocol for detection of T cell receptor (TCR) transcripts present in small amounts of synovial tissue. To expand the substrate for PCR, preamplification of cDNA was performed with a 3’ constant region primer plus either a mixture of variable region primers or random hexanucleotides. Utilizing this method improved the sensitivity of detection. This technique is applied here to the analysis of TCR transcripts in synovial biopsies from individuals with early rheumatoid arthritis (RA) and non-RA synovitis. TCR α-chain transcripts were detectable in 5/5 RA and 4/4 non-RA specimens evaluated, with β-chain transcripts detected in 4/5 early RA and 4/4 non-RA specimens evaluated. Confirmation of transcripts by sequencing of cloned PCR products verfied the specificity of amplification. The most frequently expressed TCR V region families in early RA synovitis were Vαl 1, Vαl4, Vα28, Vβ7, Vβ9 and Vβl7. Several of these V regions have previously been implicated in studies of chronic RA synovitis. Jα and Jβ region usage was similar to that seen in chronic RA, and conserved N region motifs were apparent. We conclude that it is possible to detect TCR transcripts in small synovial biopsies from individuals with early arthritis. This technique can be applied to studies of synovitis where limited quantities of synovial tissue are available.