Establishment of antigen-specific immunotherapy with artificial lymphocytes genetically engineered by antigen specific receptor and functional genes.

抗原特异性T淋巴细胞应该克隆性积累，以发挥其作用。我们已经建立了一种利用RT-PCR和T细胞受体(TCR)上的SSCP方法检测淋巴细胞群体中积累的T淋巴细胞的系统。然后，我们将我们的工作扩展到在体外重建TCR功能。利用这种方法，可以产生用于抗原特异性免疫治疗的人工淋巴细胞。存在两个技术困难。它们是来自单个T淋巴细胞的两个TCR信息的配对，以及来自接受治疗的接受者将多个基因诱导成淋巴细胞。我们已经尝试开发了几种这样的方法，并发现到目前为止，一种单细胞分选方法对配对有效。对于向淋巴细胞的基因转移，逆转录病毒载体将是最好的。我们已经成功地获得了超过50%的单基因转导。因此，超过10%的处理淋巴细胞群体可以表达3个基因。我们现在正在使用这些已建立的方法对NZB/W F1小鼠的狼疮性肾炎进行实验性免疫治疗。

Antigen-specific T lymphocytes should be clonally accumulated in order to perform their roles. We have established a system to detect such accumulated T lymphocytes in a lymphocyte population using RT-PCR and SSCP method on T cell receptor (TCR). We then extended our work to reconstitute the TCR function in vitro. With this method, artificial lymphocytes for antigen-specific immunotherapy could be generated. Two technical difficulties exist. They are the pairing of two TCR messages from a single T lymphocyte and induction of multiple genes into lymphocytes from a recipient for the therapy. We have tried to develop several of these methods and found that a single cell sorting method is so far working for the pairing. Regarding the gene transfer to lymphocytes, a retrovirus vector would be the best. We have succeeded in obtaining more than 50 % of transduction of a single gene. Therefore, more than 10 % of the treated lymphocyte population could express 3 genes altogether. We are now performing experimental immunotherapy for lupus nephritis of NZB/W F1 mice using these established methods.