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为初步探讨PKCθ在αβT细胞活化诱导细胞死亡(AICD)中的作用,采用PKCθ选择性阻断剂Rottlerin(粗糠柴毒素)预处理,抗CD3单抗再刺激αβT细胞,分别选用流式细胞术AnnexinV/PI法检测细胞早期凋亡,琼脂糖凝胶电泳检测细胞晚期凋亡。结果表明,抗CD3单抗刺激6 h后,αβT细胞早期凋亡率为28.27%±2.96%;Rottlerin(10μmol/L)预处理组,αβT细胞早期凋亡率为17.30%±2.30%。用抗CD3单抗刺激αβT细胞12 h后,出现较明显的DNA梯形条带,18 h后DNA梯形条带明显增强;用Rottlerin预处理后,DNA梯形条带明显减弱。提示PKCθ信号通路参与αβT细胞AICD的发生。
To investigate the role of PKCθ in the activation-induced cell death (AICD) induced by αβT cells, PKCθ selective blocker Rottlerin was pretreated with anti-CD3 mAb and αβT cells were stimulated with anti-CD3 mAb. Flow cytometry Cell apoptosis was detected by AnnexinV / PI assay and apoptosis was detected by agarose gel electrophoresis. The results showed that the early apoptotic rate of αβT cells was 28.27% ± 2.96% after anti-CD3 mAb stimulation for 6 h, and the early apoptosis rate of αβT cells was 17.30% ± 2.30% with Rottlerin (10 μmol / L) pretreatment. After stimulated with anti-CD3 mAb for 12 h, the more obvious DNA ladder appeared, and the DNA ladder increased significantly after 18 h. The DNA ladder decreased obviously with Rottlerin pretreatment. Tip PKCθ signal pathway involved in αβ T cell AICD.