抗CD20scFv—Fc—CD28／ξ基因修饰T细胞靶向杀伤Raji细胞的研究  

Targeted killed Raji cells by anti-CD20scFv-Fc-CD28/ξ modified T lymphocyte

作　　者：谭映霞[1] 俞康[2] 胡永仙[2] 章圣辉[1] 韩义香[1] 高中孟[1] 吴建波[1] 

机构地区：[1]温州医学院附属第一医院内科实验室,325000 [2]温州医学院附属第一医院血液室,325000

出　　处：《中华微生物学和免疫学杂志》2009年第10期919-924,共6页Chinese Journal of Microbiology and Immunology

基　　金：浙江省自然科学基金资助项目（302781,206344）

摘　　要：目的研究重组抗CD20scFv—Fc—CD28／ξ基因修饰T淋巴细胞在杀伤Raji细胞时T细胞活化情况和诱导Raji细胞凋亡的变化，进一步探讨特异基因修饰T细胞杀伤淋巴瘤细胞的机制。方法将重组质粒转染至PA317细胞中，用转染成功的PA317培养液感染外周血T淋巴细胞后，用800μg／ml的G418筛选1周后杀伤Raji细胞，分别在不同时间点用流式细胞仪检测Raji细胞Bcl-2的阳性率，用ELISA检测培养上清液中的IL-10、IFN-γ。结果Raji细胞Bcl-2的阳性率在24h内就明显下降，72h内由98．43％下降到38．45％，下降幅度明显高于对照组和空白组。72h检测IL-10发现实验组为100．3pg／ml，明显低于对照组（210．88pg／ml）和空白组（286．71pg／m1），同时IFN-γ的分泌量（1487．23pg／ml）也显著高于对照组。结论抗CD20介导的T细胞靶向杀伤Raji细胞不需要CD28的协同刺激。CD28和CD3ξ同刺激可增强T细胞抑制Raji细胞IL-10表达使其降低Bcl-2的表达从而加速靶细胞的裂解。CD28／ξ在没有外源协同刺激分子B7／CD28时能充分激活T细胞，促进其分泌IFN-γ，增强了抗CD20修饰T细胞靶向杀伤Raji细胞的能力。Objective To investigate the target killing effect on Raji cells with anti-CD20scFv-Fc- CD28/ξ gene chimeric T lymphocytes and the activation of T lymphocytes and further approach the mechanism of gene chimeric T lymphocytes target killing lymphoma cells. Methods The plasmids were transfected into retrovirus-packed PA317 cell lines by liposome. The supernatant was collected from successfully transfected PA317 cells, then co-cultured with T lymphocytes, screening with G418 800 μg/ml for 1 week. T-lymphocytes co-culture was grafted with Raji cell lines, the Bcl-2 of Raji cells were examined at timecourse by FACS. the level of IL-10 and IFN-γ was determined by ELISA. Results The Bcl-2 rate had obviously decreased within 24 h in Raji cells, the decreased amplitude was from 98.43% to 38.45% within 72 h, which is significant higher than that in control group and blank group. We found the IL-10 was 100.30 pg/ml in test group at 72 h, which was lower than that in control group （210.88 pg/ml） and blank group （286.71 pg/ml）. At the same time IFN-γ（ 1487.23 pg/ml） was higher than that of other groups. Conclusion Anti-CD20-mediated T cells target lysis Raji cells does not require CD28 costimulation. T cells can enhanced inhibited the IL-10 secretion of Raji by CD28 and CD3ξ costimulation, and also decrease the Bcl- 2 of Raji, thereby it accelerates the target cell lysis. CD28/ξ can completely activate T cell without exogenous BT/CD28 costimulation, which furthermore promote T lymphocyte secretion IFN-γ, and can also enhance the targeted killing activity of T lymphocytes to Raji cells.

关 键 词：淋巴瘤 T淋巴细胞 免疫治疗 

分 类 号：R73-3[医药卫生—肿瘤]