中文摘要：

目的:构建稳定表达人PD-L1分子的基因转染细胞株,观察其对活化的Jurkat细胞增殖和凋亡的影响。方法:将编码人PD-L1分子的全长c DNA重组入反转录病毒表达载体p EGZ-Term-R,将重组载体p EGZ-Term-R/PD-L1和辅助病毒载体用脂质体法共转染293T细胞,包装具有感染能力的完整病毒;收集含有完整重组反转录病毒的293T细胞培养上清,感染L929细胞,筛选并获得G418抗性的基因转染细胞。采用流式细胞术检测表达PDL1的基因转染细胞,命名为L929/PD-L1。采用PHA活化T细胞系来源的细胞株Jurkat,并将其与丝裂霉素预处理的L929/PD-L1细胞共培养,用细胞计数法观察L929/PD-L1细胞株对活化的Jurkat细胞增殖能力的影响,并检测其凋亡情况。结果:成功获得了稳定表达人PD-L1基因的转染细胞株L929/PD-L1。PHA可诱导Jurkat细胞活化并表达PD-1分子;L929/PD-L1与PHA刺激后的Jurkat细胞共培养,可抑制Jurkat细胞增殖,促进其凋亡。结论:成功构建了稳定表达人PD-L1的细胞株,PD-L1分子抑制活化的Jurkat细胞增殖,促进其凋亡。

英文摘要：

Objective:To construct a gene transfected cell line that stably expressing the human PD-L1 and observe its biological function on the proliferation and apoptosis of activated Jurkat cell.Methods:Human PD-L1 gene was sub-cloned into retroviral expressing vector pEGZ-Term-R.The recombinant plasmid together with its helper virus vector was cotransfected into the 293T cells.The L929 cells were infected with the supernatant of the transfected 293T cells,and then were selected with G418.The G418 resistant cells were harvested for screening PD-L1 expression by flow cytometry.L929/PD-L1 cell was cocultivated with PHA stimulated Jurkat cell.The biological effect on proliferation and apoptosis of Jurkat cell was analyzed by cell counting and flow cytometry.Results:Gene transfected line L929/PD-L1 that stably expressed the human PD-L1 was established.Membrane PD-1 expression on Jurkat cell was induced by PHA administration.L929/PD-L1 inhibited the proliferation and induced the apoptosis of activated Jurkat cell.Conclusion:Gene transfected cell line that stably expressed the human PD-L1 was established successfully.The PD-L1 on the cell line inhibited the proliferation and induced the apoptosis of activated Jurkat cell.