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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。Western blot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。
Objective: To investigate the effect of activating / inhibiting CD59 on T cell proliferation. Methods: Jurkat cells were electroporated into pSUPER-siCD59 plasmid and stimulated with CD59-activated antibody. Under the confocal laser scanning microscope, the cell turnover and the distribution and expression of CD59 on the cell membrane were observed. The cell proliferation was detected by MTT assay. Western blot was used to detect the expression of CD59 and the phosphorylation of ZAP70. Results: The transfected cells expressed green fluorescence under confocal laser scanning microscope, and the transfection efficiency was about 40%. After transfection with pSUPER-siCD59 plasmid, the fluorescence intensity of CD59 decreased, while the distribution of CD59 molecules in the cell membrane was consistent with that of normal Jurkat cells. Antibody activated CD59 in the cell membrane clustered distribution. Antibodies activated cell proliferation rate and phosphorylated ZAP70 protein expression levels were higher than the normal group (P <0.05), while the cell after electroporation plasmid is exactly the opposite. Conclusion: CD59 promotes T cell activation and proliferation through its interaction with signal transduction molecules.