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目的:研究糖基磷脂酰肌醇(GPI)锚定蛋白CD59对CD55介导T细胞信号转导的增强效应。方法:实验分为未转染的Jurkat细胞组(Ⅰ组)、转染空质粒的Jurkat细胞组(Ⅱ组)及转染CD59干扰质粒的Jurkat细胞组(Ⅲ组)。用RT-PCR检测3组细胞中的CD59基因表达水平。用噻唑蓝(MTT)比色法、免疫印迹技术和激光共聚焦扫描显微镜分别检测CD55与CD59联合作用对3组Jurkat细胞的增殖效应,Src家族酪氨酸激酶(SrcPTK)磷酸化的水平及细胞内钙离子的变化。结果:稳定转染后,Ⅲ组细胞CD59分子的表达被成功抑制。Ⅰ组和Ⅱ组细胞CD55与CD59联合作用后增殖能力,SrcPTK磷酸化水平及钙离子浓度均明显高于Ⅲ组(P<0.05);但Ⅰ组和Ⅱ组之间无差异。结论:CD59可增强CD55对T细胞信号转导的效应。
AIM: To investigate the enhanced effect of glycosylphosphatidylinositol (GPI) anchored protein CD59 on CD55-mediated T cell signaling. Methods: The experiment was divided into untransfected Jurkat cells (group Ⅰ), Jurkat cells transfected with empty plasmid (group Ⅱ) and Jurkat cells transfected with CD59 (group Ⅲ). The expression of CD59 gene in three groups of cells was detected by RT-PCR. The proliferation of Jurkat cells and the phosphorylation of Src family tyrosine kinases (SrcPTK) were detected by MTT assay, Western blotting and laser confocal scanning microscopy. Changes in calcium ions. Results: After transfection, the expression of CD59 in group Ⅲ was successfully inhibited. Proliferation, SrcPTK phosphorylation and calcium concentration in group Ⅰ and group Ⅱ were significantly higher than those in group Ⅲ (P <0.05). However, there was no difference between group Ⅰ and group Ⅱ. Conclusion: CD59 can enhance the effect of CD55 on T cell signal transduction.