TCR基因过继性免疫治疗在治疗恶性肿瘤和病毒性疾病取得了一些成功。然而，这种有效的治疗策略却存在一个亟需解决的问题，即导入的外源TCR能与内源TCR形成杂合的异二聚体（错配TCR），这种错配的TCR可能会导致自身免疫性疾病。因此有必要设法避免或减少这种错配TCR的产生。前期研究中我们利用结构域替换和三维建模方法对TCR分子进行改造，设计并构建了融合CD3ζ分子的TCRαζβζ分子，并证实该TCRαζβζ分子在T细胞表面表达增强并能避免错配。在本研究中，我们将在此基础上将该TCRαζβζ转入T细胞，对该TCRαζβζ分子在T细胞中与CD3分子的相互作用、在T细胞膜形成免疫突触的情况以及向T细胞内传递活化信号的能力进行全面的检测与分析，阐明该TCRαζβζ分子在T细胞中的信号转导通路。最后检测转TCRαζβζ基因T细胞的抗肿瘤作用，为该TCRαζβζ修饰T细胞的过继性治疗的临床应用奠定基础。

Adoptive transfer of genetically T-cell receptor (TCR) has been demonstrated the overall feasibility and clinical potential as treatments for different types of cancer. However, this promising clinical approach is limited by a serious potential consequence: the exogenous TCR mismatching with endogenous TCR chains may lead to the risk of self-reactivity. Therefore, it is urgent to reduce or avoid the generation of the mispairing TCR. In our previous study, we generated a TCRαζβζ which modified by fusing the original constant domains downstream of the extracellular cysteine of TCRα- and β- chains to complete human CD3ζ. And we had determined that the modified TCR improved expression and avoid mis-pairing on T cell surface. In this study, we will transfer the TCRαζβζ into T cells, then we will study the capacity of TCRαζβζ forming complexes with CD3 molecules. We will further characterize this TCRαζβζ in gene-engineered T cells and assess whether this receptor is able to interact with surface molecules and drive correct synapse formation in T cells. We would elucidate the signal transduction pathways of this TCRαζβζ molecule in T cells. We would elucidate the signal transduction pathways of this TCRαζβζ molecule in T cells.After that, we will test and analyze the ability of signal transduction of the TCRαζβζ. Finally, we will detect and assess the anti-tumor function of this TCR gene-engineered T cells in vitro.Taken together, this study may explain the anti-tumor effect and its molecular mechanism of this TCRαζβζ. Taken together, this study may provide further understanding of the TCRαζβζ in adoptive transfer of TCR gene therapy.