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目的扩增弥漫大B细胞淋巴瘤(diffuse large B cell lymphoma,DLBCL)相关抗原特异T细胞克隆的全长TCRα和β链基因序列,并对其三维空间结构进行预测。方法从已鉴定的2例伴有克隆性增殖TCR Vα6和Vβ13亚家族T细胞的DLBCL患者外周血中克隆全长TCRα和β基因序列,利用Primer Premier 5.0分析软件推算其相应的氨基酸序列,并通过自动化蛋白质模建服务器SWISS-MODEL进行三维空间结构预测。然后利用蛋白质三维图像分析软件Rasmol对所获得的TCR三维空间结构进行显示和分析。结果获得DLBCL相关抗原特异TCRα6和β13全长基因序列及其氨基酸序列。2例患者的TCR Vα6的空间结构与PDB ID:3ffcD模型相似,序列同源性分别为92.16%和91.71%;而2个TCR Vβ13也均具有同一个空间结构模型PDB ID:2ialD,序列同源性分别为92.95%和92.18%。结论来自不同DLBCL患者外周血中克隆性增殖的TCR Vα6或Vβ13亚家族T细胞的TCR具有高度同源性的空间结构,提示其可能识别相同的肿瘤抗原表位。
OBJECTIVE: To amplify the full-length TCRα and β chain genes of diffuse large B cell lymphoma (DLBCL) -associated antigen-specific T cell clones and to predict their three-dimensional spatial structure. Methods The full-length TCRα and β gene sequences were cloned from peripheral blood of two DLBCL patients with clonal proliferative TCR Vα6 and Vβ13 subfamily T cells. Primer Premier 5.0 software was used to deduce the corresponding amino acid sequence and to pass Automated protein modeling server SWISS-MODEL 3D spatial structure prediction. Then the three-dimensional structure of TCR obtained was displayed and analyzed by Rasmo protein three-dimensional image analysis software. Results The full-length gene sequences of DLBCL-associated antigen-specific TCRα6 and β13 and their amino acid sequences were obtained. The spatial structure of TCR Vα6 in two patients was similar to that of PDB ID: 3ffcD, and their sequence homologies were 92.16% and 91.71% respectively. The two TCR Vβ13 also shared the same spatial structure model PDB ID: 2ialD, Sexuality was 92.95% and 92.18% respectively. Conclusion The TCRs of TCR Vα6 or Vβ13 subfamily T cells from clonal proliferation in peripheral blood of patients with different DLBCL have a highly homologous spatial structure suggesting that they may recognize the same tumor epitope.