死亡受体5诱导Jurkat细胞凋亡的作用  

Role of DR5 in TRAIL induced apoptosis of Jurkat cells

作　　者：白慧玲[1] 龚非力[1] 马远方[2] 

机构地区：[1]华中科技大学同济医学院免疫室,湖北武汉430030 [2]河南大学医学院细胞与分子免疫学实验室,河南开封475001

出　　处：《第四军医大学学报》2005年第22期2068-2071,共4页Journal of the Fourth Military Medical University

基　　金：河南省医学科技创新人才工程项目(2002-119)

摘　　要：目的:探讨DR5在TRAIL诱导Jurkat细胞凋亡中的作用.方法:用含人全长DR5细胞外全长结构域重组DR5免疫BALB/c小鼠,制备抗DR5 mAb.将96孔U型细胞培养板加1×105/孔Jurkat细胞和5 mg/L抗DR5 mAb,采用TRAIL试剂盒检测细胞凋亡率.结果:抗DR5 mAb与Jurkat细胞表面的DR5作用后,TRAIL对Jurkat细胞的杀伤功能几乎被抗DR5 mAb完全阻断,阻断率高达90.49%.结论:DR5在TRAL诱导的Jurkat细胞凋亡中起关键作用.AIM： To study the role of DR5 in TRAIL apoptotic signal transduction in Jurkat cells. METHODS： cDNA containing full-length extracellular domain of human DR5 was cloned into pGAPZa. Recombinant Pichia pastoris clone generated via homologous recombination secreted high levels of sDRS. BALB/c mice were immunized with sDR5 in CFA to prepare anti-DR5 mAb. The binding of anti-DR5 mAb was measured for surface expression of TRAIL receptor by flow cytometric analysis. Jurkat cells were tested for their susceptibility to apoptosis induced by TRAIL with TRAIL apoptosis kit so as to study the correlation between DR5 expression level and TRAIL sensitivity. The level of DR5 expression on Jurkat cell line was examined by flow cytometry. The rates of TRAIL-induced apoptosis and anti-DR5 mAb blocking on Jurkat cells were tested by flow cytometry with TRAIL apoptosis kit. RESULTS： The killing role of TRAIL was blocked in Jurkat cells pretreated with anti-DR5 mAb. The average percentage of blocking was 90.49%. CONCLUSION ： Anti-DR5 mAb can specifically bind to DR5 and DR5 is expressed at high levels on Jurkat cells. DR5 plays a very key role in TRAIL induced apoptosis in Jurkat cells. DR5 expression is important for the induction of apoptosis by TRAIL.

关 键 词：JURKAT细胞 TRAIL 细胞凋亡 DR5 抗体 单克隆 

分 类 号：R320.84[医药卫生—人体解剖和组织胚胎学]