不同等位基因E7/HLA-DR四聚体结合的结核患者CD4+T细胞CDR3区序列分析
Analysis of TCR CDR3 sequences of E7/HLA-DR tetramers-bound CD4+ T cells from tuberculosis patients by using tetramers constructed with different HLA-DRB1 alleles
摘要目的 探讨机体抗结核免疫过程中CD4+T细胞TCR与多肽/MHC之间的识别和结合规律.方法 从9例结核患者胸水中用等位基因不同的结核多肽E7/HLA-DR四聚体磁珠分选出E7结合阳性CD4+T细胞,提取其RNA逆转录成cDNA作为模板扩增TCRα链和β链CDR3区核酸片段,并比较其长度和序列特征.结果 等位基因不同的E7/HLA-DR四聚体能够识别同一患者CDR3区序列相同或结构功能相似的CD4+T细胞克隆.结论 结核患者体内CD4+T细胞识别和结合抗原多肽时有一定的HLA-DR限制性,但是主要以多肽特异性为主,这为进一步探讨机体抗结核免疫中CD4+T细胞和MHC之间的抗原提呈机制提供了数据.
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abstractsObjective To investigate the recognition and binding mechanism between TCR of CD4+ T cells and peptide/MHC ( major histocompatibility complex) in immune responses against Mycobacte-rium tuberculosis (MTB). Methods Nine patients with tuberculosis (TB) were recruited in the present study. Peptide E7-bound CD4+T cells were sorted by using E7/HLA-DR tetramers constructed with different HLA-DRB1 alleles from pleural fluid ( PLF) of TB patients. Total RNA was extracted and reversely tran-scribed into cDNA, which was used as the template to amplify complementarity determining region 3 (CDR3) fragments of T cell receptor (TCR) α and β chains. Amino acid sequences, spectratypes and lengths of the amplified CDR3 fragments were analyzed. Results The CDR3 amino acid sequences of E7-bound CD4+ T cells were identical completely or partially in a single individual. E7/HLA-DR tetramers with different HLA-DRB1 alleles were capable of recognizing and binding CDR3 fragments with identical sequence or similar structure and function in one single individual. Conclusion The TCRαandβchains CDR3 line-age of CD4+ T cells of TB patients apparently drifts, and the predominant CDR3 sequences of TCRαandβchains that recognize MTB antigen exhibit peptide specificity with some certain HLA-DR restriction. This study illustrates the possible causes and mechanisms of peptide-specific CD4+T cells related antigen presen-tation against MTB.
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