Anti-CD20scFv/IgGFc/CD80/CD28/zeta转染人T淋巴细胞对原代CLL细胞的作用

Anti-CD20scFv/CD80/CD28/zeta specific T lymphocytes eradicate primary chronic lymphocytic leukemia cells in vitro

目的:将已成功构建表达anti-CD20scFv/CD80/CD28/zeta转染人T淋巴细胞,体外观察该类细胞特异性清除CD20+原代慢性淋巴细胞白血病(CLL)细胞的能力,为肿瘤的过继免疫治疗提供新思路。方法:将本室成功构建的含anti-CD20scFv/IgGFc/CD80片段的PLNCX质粒,转染PA317包装细胞,挑取高滴度的包装细胞株收获逆转录病毒,用收获的病毒感染刺激分裂的人外周血T细胞,经G418筛选后与CD20+原代CLL细胞在体外共同培养,在显微镜下观察CD20+的原代CLL细胞生长状态,ELISA检测试剂盒检测T细胞分泌细胞因子的功能。结果:重组基因修饰的T细胞能在体外杀伤CD20+原代CLL细胞,而对CD20-细胞无杀伤作用;同时靶细胞为CD20+组上清液中IL-2(1301.00pg/ml)和IFN-γ(602.18pg/ml)水平与CD20-组相比明显升高。结论:嵌合锚定T细胞能够成功构建;该类T细胞在体外能特异性杀伤CD20+的原代CLL细胞。

Objective：To construct anti-CD20scFv/CD80/CD28/zeta recombinant gene modified T cells,test its effectiveness of eradicating CD20 positive primary chronic lymphocytic leukemia （CLL） cells and provide a promising tool for tumor adoptive immunotherapy. Me-thods： The recombinant vectors were transduced into PA 317 cells and high titer retroviruses were obtained to infect human peripheral blood T lymphocytes. Resistant T cells were obtained by G418 selection for one week. Then transduced T lymphocytes and primary CLL cells were cocultured. The status of primary chronic lymphocytic leukemia cells were observed by microscope. The level of IL-2 and IFN-γ in the culture medium were measured. Results： Primary T cells expressing anti-CD20scFv/IgGFc/CD80/CD28/zeta could be constructed successfully. These T cells were able to lyse CD20＋ targets and secrete high levels of IL-2（1301.00 pg/ml） and IFN-γ（602.18 pg/ml） in vitro. Conclusion： ①Recombinant gene modified T cells can be constructed successfully. ②Recombinant gene modified T cells can specially kill CD20 positive primary CLL cells in vitro.