活化／抑制CD59对T淋巴细胞增殖的影响

目的：研究活化／抑制cD59分子对T细胞增殖的影响。方法：Jurkat细胞分别电转入pSUPER-siCD59质粒及用CD59活化抗体刺激。激光共聚焦显微镜下观察细胞的电转情况及cD59分子在细胞膜上的分布及表达；MTT比色法检测细胞的增殖。Westernblot检测CD59分子表达及T细胞活化相关蛋白ZAP70磷酸化水平。结果：激光共聚焦显微镜下可见电转染细胞表达绿色荧光，转染效率约为40％。转染pSUPER-siCD59质粒后CD59荧光强度强度降低，CD59分子均匀分布于细胞膜与正常Jurkat细胞分布一致。抗体活化后CD59在细胞膜成簇状分布。抗体活后细胞增殖速率和磷酸化ZAP70的蛋白表达水平均高于正常组（P〈0．05），而细胞电转质粒后则恰恰相反。结论：CD59通过与信号转导分子的相互作用促进T细胞活化增殖。

The Effect of Activation / down-regulation of CD59 on T Lymphocyte Proliferation

Objective： The present study aims to investigate the effect of CD59 on T lymphocyte proliferation. Methods： Jurkat cells were transfected with pSUPER-siCD59 plasmids by electroporetion or stimulated by anti-CD59 antibody. Confocal laser scanning microscopy was used to observe the transfection efficiency and cellular location of immunostrained CD59. The cells proliferation was measured by MTT assay. Furthermore, western blot was performed for the detection of CD59 expression and ZAP70 phosphorylation. Results： 40% of Jurkat cells were successfully transfected and showed lower fluorescence staining. In contrast to the uniform distribution, CD59 formed clusters on the cell surface upon stimulation with anti-CD59 antibody. And also compared with the untreated control cells, anti-CD59 stimulated-cells showed a higher level of proliferation and ZAP70 phosphorylation, while knocking down of CD59 led to the decreased cell proliferation and ZAP70 pho-sphorylation. Conclusion： CD59 has a positive effect on T cell activation and proliferation.