DR5在TRAIL诱导Jurkat细胞凋亡中的作用

The role of DR5 in TRAIL-induced apoptosis for Jurkat cell line

目的 :研究DR5在介导TRAIL凋亡信号中的作用。方法 :用含人DR5细胞外全长结构域重组DR5免疫BALB C小鼠 ,制备抗DR5单克隆抗体 ;流式细胞仪检测Jurkat细胞表面DR5表达水平 ;采用TRAIL凋亡检测试剂盒 ,检测Jurkat细胞凋亡率及抗DR5单克隆抗体对TRAIL诱导细胞凋亡的阻断率。结果 :DR5在Jurkat细胞表面的表达率为 94 83% ,TRAIL和抗TRAIL单克隆抗体能够诱导Jurkat细胞凋亡 ,呈现明显的剂量相关性 ,TRAIL浓度在 5 0～ 10 0ng ml时 ,杀伤率达 90 %以上。预先用抗DR5单克隆抗体与Jurkat细胞作用后 ,TRAIL对Jurkat细胞的杀伤功能几乎完全被mAb所阻断 ,其平均阻断率达90 4 9%。结论 :DR5在TRAIL诱导Jurkat细胞凋亡中起着十分关键的作用。

Objective:To study the role of DR5 in TRAIL apoptotic signal transduction. Methods:BALB/C mice were immunized with recombinant DR5 that contains the full-length extracellular domain of the human DR5. Anti-DR5 mAb was generated by hybridoma. 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 percentage of DR5 expression on Jurkat cells was 94 83%. TRAIL and anti-TRAIL mAb could kill Jurkat cells on dose-dependent, and the killing rate was 90% in the concentration of 50～100 ng/ml. The killing role of TRAIL could be blocked on Jurkat cells pretreated with anti-DR5 mAb. The average percentage of blocking was 90 49%.Conclusion:DR5 plays a very key role in TRAIL induced apoptosis in Jurkat cells.