| 李园园 高美华 丛蓓蓓 王丽娜 王冰 张蓓 李营 梁洁.活化/抑制CD59 对T 淋巴细胞增殖的影响[J].,2014,14(19):3631-3634 |
| 活化/抑制CD59 对T 淋巴细胞增殖的影响 |
| The Effect of Activation / down-regulationof CD59 on T Lymphocyte Proliferation |
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| DOI： |
| 中文关键词: siCD59 电转染 Jurkat 磷酸化蛋白 |
| 英文关键词: SiCD59 Electric transfection Jurkat Phosphorylation |
| 基金项目:国家自然科学基金项目（81273206） |
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| 中文摘要: |
| 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。 |
| 英文摘要: |
| 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 uniformdistribution,
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. |
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