结直肠癌T细胞克隆TCRβ基因CDR3区氨基酸序列的初步分析

Sequence Analysis of the TCRβ CDR3 Region of T Cell Clones in Colorectal Cancer

摘要: 目的　通过建立结直肠癌肿瘤浸润性淋巴细胞 (tumorinfiltratinglymphocytes, TIL)和术前外周血 (peripheralbloodlymphocytes, PBL)中抗肿瘤T细胞克隆的TCRβ基因谱型, 确定抗肿瘤T细胞克隆的分子特征。方法　采用RT PCR方法扩增结直肠癌TIL和PBL的TCRβ基因的 2 4个不同V家族基因片段, 经过测序凝胶分离和特殊银染色, 结合测序结果确定TCRβ基因谱型。结果　结直肠癌患者具有TIL和 /或PBL中T淋巴细胞的克隆性增殖。有些T细胞克隆表达相同的CDR3区氨基酸基序。CDR3区不同位点具有特征性的疏水性和极性氨基酸以及甘氨酸的分布。结论　通过分析结直肠癌抗肿瘤T细胞克隆特征性的CDR3区氨基酸序列, 将有益于发现新的抗肿瘤免疫治疗方法。

Abstract: Objective To study the molecular characteristics of anti-tumor T cell clones by analyzing the sequence of TCRβ CDR3 region from tumor-infiltrating lymphocytes (TIL) and peripheral blood lymphocytes (PBL) of patients with colorectal cancer. Methods The 24 TCR(V gene from TIL and PBL in pre-operation and post-operation were amplified by the method of combined reverse transcriptase-polymerase chain reaction (RT-PCR) with denaturation polyacrylamide sequencing gel eletrophoresis and gene sequencing,Which is ca...