Detection of T cell receptors in early rheumatoid arthritis synovial tissue.
Detection of T cell receptors in early rheumatoid arthritis synovial tissue.
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早期类风湿性关节炎滑膜组织中 T 细胞受体的检测。
DOI：
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发表时间：
1995
期刊：
影响因子：
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通讯作者：
William V. Williams
中科院分区：
文献类型：
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作者：
Thaila Ramanujam;Monica Luchi;Ralph Schumacher;Samuel Zwillich;Chang Pei Chang;Peter E. Callegari;Joan M. Von Feldt;Q. Fang;David B. Weiner;William V. Williams
早期滑膜炎患者很少有滑膜组织，但滑膜活检除外。然而，T细胞群在滑膜炎的发展早期可能富含抗原特异性细胞，并对疾病的发病机制至关重要。为了研究早期滑膜炎中的T细胞库，我们利用PCR方案检测少量滑膜组织中存在的T细胞受体（TCR）转录本。为了扩增用于PCR的底物，用3'恒定区引物加上可变区引物的混合物或随机六核苷酸进行cDNA的预扩增。利用该方法提高了检测的灵敏度。该技术在此应用于分析早期类风湿性关节炎（RA）和非RA滑膜炎个体滑膜活检中的TCR转录本。在5/5例RA和4/4例非RA样本中检测到TCR α链转录物，在4/5例早期RA和4/4例非RA样本中检测到β链转录物。通过克隆PCR产物测序确认转录本，验证了扩增的特异性。在早期RA滑膜炎中最常表达的TCR V区家族是V α 11、V α 14、V α 28、V β 7、V β 9和V β 17。这些V区中的几个先前已涉及慢性RA滑膜炎的研究。J α和J β区域的使用与慢性RA中观察到的相似，并且保守的N区域基序很明显。我们的结论是，这是可能的检测TCR转录小滑膜活检的个人早期关节炎。（250字处删节）
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 alpha-chain transcripts were detectable in 5/5 RA and 4/4 non-RA specimens evaluated, with beta-chain transcripts detected in 4/5 early RA and 4/4 non-RA specimens evaluated. Confirmation of transcripts by sequencing of cloned PCR products verified the specificity of amplification. The most frequently expressed TCR V region families in early RA synovitis were V alpha 11, V alpha 14, V alpha 28, V beta 7, V beta 9 and V beta 17. Several of these V regions have previously been implicated in studies of chronic RA synovitis. J alpha and J beta 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.(ABSTRACT TRUNCATED AT 250 WORDS)