Yin Qingsong,Tan Huo,Chen Shaohua,Yang Lijian,Li Bo,Ye Jingmei,Li Yangqiu.Cloning of antigen-specific TCR α and β genes and their 3-D structure prediction[J].J Amry Med Univ (J Third Mil Med Univ),2011,33(04):349-352.
Yin Qingsong,Tan Huo,Chen Shaohua,Yang Lijian,Li Bo,Ye Jingmei,Li Yangqiu.Cloning of antigen-specific TCR α and β genes and their 3-D structure prediction[J].J Amry Med Univ (J Third Mil Med Univ),2011,33(04):349-352.DOI:
Cloning of antigen-specific TCR α and β genes and their 3-D structure prediction
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TCR α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TCR α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TCR α6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。
Abstract
Objective To amplify the full-length sequences of diffuse large B cell lymphoma (DLBCL)-associated antigen-specific TCR α and β genes and to predict three-dimensional (3-D) structure of the proteins. Methods The full-length TCR genes were amplified from the peripheral blood mononuclear cells (PBMCs) derived from 2 pathologically identified DLBCL patients for the TCR Vα6 and Vβ13 subfamily T-cell clones
and their amino acid sequences were deduced by primer premier 5.0 analytical software. 3-D spatial structures of the TCR α and β were modeled by SWISS-MODEL
then displayed and analyzed by the analytical software of Rasmol. Results The full-length TCR genes and their amino acid sequences were obtained successfully. the TCR Vα6 chains from the 2 cases of DLBCL patients were of high homology not only in sequence (92.16% and 91.71%
respectively)
but also in the 3-D spatial structures
which displayed the same model (PDB ID:3ffcD) by the SWISS-MODEL homology-modeling server. While the TCR Vβ13 chains also used an identical model (PDB ID:3dx9D)
and sequence homology was 92.95% and 92.18%
respectively. Conclusion The TCR of Vα6 or Vβ13 subfamily T cell clones have high homology in the 3-D spatial structures from the PBMCs of 2 cases of DLBCL patients
suggesting that they might recognize similar tumor associated antigen epitope.