肿瘤源性热休克蛋白gp96诱导淋巴细胞反应及其抗肿瘤效应  

Tumor-derived HSP gp96 induces T cell responses and its significance in tumor immunity

作　　者：朱昌来[1] 林琳[1] 张天一[1] 

机构地区：[1]南通大学神经再生重点实验室,南通226001

出　　处：《中国免疫学杂志》2007年第3期234-237,共4页Chinese Journal of Immunology

基　　金：江苏省教育厅资金(03KJD320183)资助

摘　　要：目的研究肿瘤细胞来源的热休克蛋白gp96-多肽复合物在体外诱导脾淋巴细胞的特异性细胞毒性T淋巴细胞(CTL)反应。方法利用蛋白纯化技术、SDS-PAGE凝胶电泳及Western blot法分离纯化、鉴定gp96-多肽;通过流式细胞术、免疫荧光技术、CCK-8法等检测经gp96多肽诱导的CD8+T细胞及其抗肿瘤效应。结果经鉴定获得纯化的热休克蛋白;流式细胞仪检测表明,经gp96-肽复合物诱导后的CD8+T细胞比例达到近70%,远远高于对照组的35%、26%;该活化的CTL细胞在效靶比为50∶1时的肿瘤杀伤率达72%,与对照组相比具有统计学意义;激光共聚焦显微镜观察证实实验诱导组的培养上清能诱导H22肿瘤细胞凋亡的形态学改变。结论肿瘤来源的热休克蛋白gp96-肽复合物能诱导小鼠脾淋巴细胞的CTL反应,该活化的CTL具有特异性抗H22肿瘤细胞的免疫作用,并能分泌免疫活性物质诱导H22肿瘤细胞凋亡。Objective：To explore the availability for tumor-derived gp96 to induce specific CTL activity of splenocytes in vitro. Methods：gp96 purified by the techniques for protein extraction and identified by SDS-PAGE gel electrophoresis and Western blot method ; CD8^＋T cell induced by gp96 and CTL activity detected by flow cytometry, immunofluorescence technic and CCK-8 assay. Results： gp96 was identified by SDS-PAGE and Western blot ; Analysis with FCM showed that the number of CD8^＋T cells （ nearly 70% ） was obviously increased after pulsed with gp96-peptide complexes as compared with that of control groups （35%, 26%, P 〈 0. 05 ） ; The killing rate of CTL on target cells（ H22 tumor cells） was 72% at the ratio of E（Effect） to T（Target） as 50： 1, which have the significant difference compared with the control groups. The morphologic change of apoptosis of H22 tumor cells after treated by culture supermatant of experimental group can be observed with LSCM. Conclusion：H22 tumor cells derived gp96 peptide complexes can induce CTL response of spleen lymphocyte of mouse in virto ; Besides, those CTLs show the specific anti-tumor effect to H22 tumor cells and screat the immunologic active material to induce apoptosis of H22 tumor cells.

关 键 词：GP96-肽复合物 H22细胞 细胞毒性T淋巴细胞 细胞凋亡 

分 类 号：R735[医药卫生—肿瘤]