SARS-CoV X4基因转染对T细胞TNF-α及IL-6分泌的影响

Study on cytokine secretion of T cell after SARS-CoV X4 gene transfection

目的:通过检测SARS-CoV X4基因转染后T细胞细胞因子分泌的变化,研究SARS-CoV X4蛋白潜在的功能及其作用机制,并探讨其与SARS-CoV致病机制的关系。方法:利用RosetteSep分离法分离外周血T细胞,以Amaxa核转染仪将pEGFP-SARS-CoV X4真核表达载体转入T细胞,应用激光共聚焦显微镜观察其瞬时表达情况,流式细胞仪检测转染效率,CBA技术检测T细胞细胞因子。结果:SARS-CoV X4基因转染后12h得到了明显表达,共聚焦显微镜观察到EGFP绿色荧光表达,流式细胞仪检测其表达率为245;与空载体转染组相比,SARS-CoV X4基因转染能显著促进T细胞IL-6及TNF-α的分泌。结论:SARS-CoV X4蛋白可诱导T细胞分泌前炎症因子IL-6及TNF-α,SARS-CoV X4蛋白可能在SARS发病机制中起重要的作用。

Objective： To explore the function of the SARS - CoV X4 protein and its role in SARS pathogenesis, by detecting the change of cytokine secretion of T cells after transfection of SARS - CoV X4 gene. Methods： T cells were transfected with pEGFP - SARS - CoV ）（4 expression vector by Amaxa Nucleofector , and the green fluorescence of EGFP were detected with flow cytometry and confocal microscopy; The production of cytokine were investigated by Cytometry Beads Assay. Results： The green fluorescence of EGFP could be detected by with flow cytometry, confocal microscopy; the secretion of IL- 6 and TINT - α increased remarkably in SARS - CoV X4 transfection group. Conclusion： SARS - CoV X4 can promote T cells to secrete preinflammatory eytokine IL- 6 and TNF- α; it may play an important role in the pathogenesis of SARS.