Yao Xin Sheng, Diao Ying, Sun Wan Bang, Luo Jun Min, Qin Ming, Tang Xian Ying
Department of Immunology, Zunyi Medical College, Zunyi, China.
Cell Mol Immunol. 2007 Jun;4(3):215-20.
Analysis of complementarity determining region 3 (CDR3) length of T lymphocyte receptors (TCRs) by immunoscope spectratyping technique has been used successfully to investigate the diversity of TCR in autoimmune diseases and infection diseases. In this study, we investigated the patterns of CDR3 length distribution for all 32 TCR AV gene families in human peripheral blood lymphocytes of four normal volunteers by the immunoscope spectratyping technique. It was found that PCR products exhibited an obscure band on 1.5% agarose gel electrophoresis. Each TCR AV family exhibited more than 8 bands on 6% sequencing gel electrophoresis. The CDR3 spectratyping of all TCR AV families showed a standard Gaussian distribution with different CDR3 length, and the expression frequency of CDR3 was similar among the gene families. Most of CDR3 in TCR AV family recombine in frame. However, some of the CDR3 showed out-of frame gene rearrangement. Additionally, we found that in some of TCR AV families there were 18 amino acid discrepancies between the longest CDR3 and shortest CDR3. These results may be helpful to further study the recombination mechanism of human TCR genes, the TCR CDR3 gene repertoire, and the repertoire drift in health people and disease state.
利用免疫谱型分析技术分析T淋巴细胞受体（TCR）互补决定区3（CDR3）的长度，已成功用于研究自身免疫性疾病和感染性疾病中TCR的多样性。在本研究中，我们采用免疫谱型分析技术，对4名正常志愿者外周血淋巴细胞中所有32个TCR AV基因家族的CDR3长度分布模式进行了研究。结果发现，PCR产物在1.5%琼脂糖凝胶电泳上呈现模糊条带。每个TCR AV家族在6%测序凝胶电泳上呈现8条以上条带。所有TCR AV家族的CDR3谱型分析显示，不同CDR3长度呈标准高斯分布，且各基因家族中CDR3的表达频率相似。TCR AV家族中的大多数CDR3以框内方式重组。然而，部分CDR3显示出框外基因重排。此外，我们发现部分TCR AV家族中最长CDR3与最短CDR3之间存在18个氨基酸差异。这些结果可能有助于进一步研究人类TCR基因的重组机制、TCR CDR3基因库以及健康人和疾病状态下的库漂移。
