Analysis of the CDR3 length repertoire and the diversity of TCR alpha chain in human peripheral blood T lymphocytes.
Analysis of the CDR3 length repertoire and the diversity of TCR alpha chain in human peripheral blood T lymphocytes.
复制标题
人外周血T淋巴细胞中CDR3长度库和TCRα链多样性分析。
作者：
免疫荧光光谱技术分析T淋巴细胞受体（TCR）互补决定区3（CDR3）的长度已成功地用于研究自身免疫性疾病和感染性疾病中TCR的多样性。本研究应用免疫组化技术对4名正常人外周血淋巴细胞32个TCR AV基因家族的CDR3长度分布进行了研究。PCR产物在1.5%琼脂糖凝胶电泳上显示一条模糊带。每个TCR AV家族在6%测序凝胶电泳上显示8条以上的条带。所有TCR AV家族的CDR3谱均呈标准的高斯分布，但CDR3长度不同，且各基因家族间CDR3的表达频率相似。TCR AV家族中的CDR3大部分在框内重组。然而，一些CDR3显示出框外基因重排。此外，我们发现在一些TCR AV家族中，最长的CDR 3和最短的CDR 3之间存在18个氨基酸的差异。这些结果有助于进一步研究人TCR基因的重组机制、TCR CDR3基因库以及TCR CDR3基因库在健康人和疾病状态下的漂移。
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.
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