抗人DR5单克隆抗体(mDRA-6)对Jurkat细胞作用研究

目的:观察抗肿瘤坏死因子相关凋亡诱导配体(TRAIL)的死亡受体5(DR5)单克隆抗体—mDRA-6对Jurkat细胞的凋亡作用。方法:制备抗人DR5单抗-mDRA-6;检测Jurkat细胞表面DR5表达率;荧光显微镜下观察mDRA-6作用下Jurkat细胞形态变化;MTT法计算mDRA-6对Jurkat细胞存活的影响;FITC-AnnexinⅤ及PI双染流式细胞仪检测mDRA-6对Jurkat细胞凋亡率影响;琼脂糖凝胶电泳检测mDRA-6对Jurkat细胞DNA片段化的作用。结果:Jurkat细胞表面DR5表达率为94.8%;mDRA-6使Jurkat细胞染色质边集、断裂,细胞出芽,凋亡小体形成;MTT法显示mDRA-6具有明显的Jurkat细胞杀伤作用,1.563 mg/L的mDRA-6可使Jurkat细胞死亡60.50%;AnnexinⅤ及PI双染显示0.3mg/L的mDRA-6作用10 h,Jurkat细胞凋亡率达43.22%;10 mg/L mDRA-6作用HL-60细胞3h,DNA琼脂糖凝胶电泳显示明显的“梯形”条带。结论:抗DR5单抗-mDRA-6能够诱导Jurkat细胞凋亡。

Effects of anti-human DR5 monoclonalantibody(mDRA-6) on Jurkat cell

Objective:To observe apoptotic effect of anti-human DR5(death receptor 5 of TRAIL) monoclonal antibody(mDRA-6) on Jurkat cell apoptosis.Methods: monoclonal antibody(mDRA-6) was made;the level of DR5 expression on Jurkat cells was detected;the morphologic changes of Jurkat cells were observed by a microscope;apoptotic effects of mDRA-6 on Jurkat cells were detected by MTT and FCM analysis,DNA fragmentation were detected by agarose gel electrophoresis.Results: DR5 expression on Jurkat cells was 94.8%;Chromatic margination,budding and apoptotic bodies were observed in Jurkat cells treated by mDRA-6;aiability of Jurkat cells is decreseased and apoptosis of Jurkat cells increased obviously,"DNA ladder" in mDRA-6-treated Jurkat cells was exhibited on agarose gel electrophoresis.Conclusion: Apoptosis of Jurkat cell can be induced by anti-human DR5 monoclonal antibody(mDRA-6)