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目的 研究HIV 1感染中相关细胞因子IFN γ对人类疱疹病毒 8型 (HHV 8)Rta基因启动子活性影响 ;评价Rta基因启动子在多种细胞中的启动活性。方法 将已构建的HHV 8Rta启动子 +虫荧光素酶报告基因重组质粒和含HIV 1Tat基因重组表达质粒转染多种细胞 ,转染后 2 4h加入IFN γ和 或重组HIV 1gp12 0刺激 ,刺激后 2 4h收集细胞 ,进行虫荧光素酶活性检测。试验同时以佛波酯类化合物TPA刺激为阳性对照 ,进行最佳刺激时间点的选择和不同转染方法转染效率的比较。结果 ①HHV 8Rta启动子启动活性在TPA刺激后的 2 4h达到最高峰 ;②HHV 8Rta启动子启动活性的发挥不依赖于HHV 8和 或EBV基因组的存在 ,且在血管内皮细胞 (HUVECs)中启动活性最佳 ;③IFN γ可以诱导Rta启动子活性 ;但HIV 1Tat和gp12 0蛋白与IFN γ协同上调Rta启动子活性的能力较弱。结论 HIV 1感染中相关细胞因子IFN γ至少部分通过Rta基因启动子激活HHV 8的复制
Objective To investigate the effect of cytokine IFNγ on the promoter activity of human herpesvirus 8 (HHV 8) Rta gene in HIV-1 infection and to evaluate the promoter activity of Rta gene promoter in various cells. Methods The recombinant plasmids of HHV 8Rta promoter + luciferase reporter gene and HIV 1Tat gene recombinant plasmids were transfected into a variety of cells. After stimulated with IFNγ and recombinant HIV 1gp12 0 24 h after transfection, 2 4h cells were collected for luciferase activity detection. At the same time, the TPA stimulated by phorbol ester compound was used as positive control, and the optimal stimulation time point was selected and compared with the transfection efficiency of different transfection methods. Results ① The promoter activity of HHV 8Rta reached its peak at 24 h after TPA stimulation. ② The initiation activity of HHV 8 Rta promoter did not depend on the presence of HHV 8 and / or EBV genome and was most active in endothelial cells (HUVECs) (3) IFNγ can induce Rta promoter activity; however, the ability of HIV 1Tat and gp12 0 proteins to co-upregulate Rta promoter activity with IFNγ is weak. Conclusion The related cytokine IFNγ in HIV-1 infection activates the replication of HHV 8 at least partially through the Rta gene promoter