摘要
目的研究IBP表达抑制后对T细胞凋亡产生的影响。方法构建IBP siRNA稳定表达及阴性对照载体,脂质体法转染Jurkat T细胞,G418筛选稳定转染细胞株,用anti-CD3和CD28 mAb刺激介导的TCR信号方式作用于稳定转染细胞后,以3H-TdR掺入实验检测细胞增殖,Annexin-v/PI双参数法经流式细胞仪检测细胞凋亡。结果TCR信号刺激下,阴性对照组细胞的3H-TdR掺入量在24 h时下降为未刺激对照组的67%,随时间延长其下降更显著,且24 h发生明显凋亡,凋亡率为21.96%;IBP siRNA表达载体转染细胞在不同时相点的增殖均不受影响,24 h凋亡率仅为2.12%,比阴性对照组细胞显著降低。结论IBP缺失能使TCR信号刺激下的细胞激活后凋亡受到抑制,提示IBP可能参与T细胞免疫自稳状态的调节。
Objective To study the effects of the depressed expression of IBP on T cell apoptosis. Methods The stable IBP siRNA and negative control vectors were established, and then transfected into Jurkat T cells by means of hposome＇ s method. The stable transfected cells were screened by G418, and then stimulated with anti-CD3 and CD28 mAb. The proliferation and apoptosis of the stimulated cells were evaluated by ^3 H-TdR incorporation and Annexin-v/PI staining, respectively. Results After stimulation of TCR signals, the ^3 H-TdR incorporation of negative control cells decreased to 67% against unstimulated control cells and apoptosis ratio raised to 21.96% at 24 h; while the proliferation of cells transfected with IBP siRNA vector had no variation compared with unstimulated control cells, and the apoptosis ratio of the transfected cells was only 2.12 % at 24 h, much lower than negative control cells. Conclusion Loss of IBP could inhibit normal apoptosis in reactivated cells stimulated by TCR signals, which indicated that IBP may participate in regulating immunological homeostasis.
出处
《免疫学杂志》
CAS
CSCD
北大核心
2007年第3期248-251,共4页
Immunological Journal
基金
国家自然科学基金资助项目(30471577)
