【摘要】 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的复制更多还原

【Abstract】 Objective To investigate effect of interferon ga mma(IFN-γ) being similar to that produced by HIV-1-infected T cells on Rta promoter activity of human herpesvirus 8(HHV-8) and to evaluate its promotin g activity. Methods Transfection of cells with HHV-8 Rta promoter-driven luciferase construct which was previously built and recombinan t expression plasmid named pEV containing HIV-1 Tat encoding gene was performed and stimulation of transfected cells with 12-O-tetradecanoylphorbol-13-acet ate(TPA) and HIV-1 gp120 was carried out at 24 hours after transfection to in duce luciferase gene expression. Then, calculation of the relative luciferase ac tivity unit(RLU) was conducted 24 hours after stimulation. Meanwhile, comparis on of transfection efficiency by different transfection methods and choice of op timal time range of Rta promoter activity were performed with TPA treatment as p ositive control. Results HHV-8 Rta promoter activity rea ched the peak 24 hours after transfection and was not dependent on intact struct ure of HHV-8 and/or Epstein-Barr virus(EBV) genome present in cells. Particu larly high level of induction of Rta promoter activity appeared to be statistica lly significant in human umbilical veil endothelial cells(HUVECs). Recombinant IFN-γ upregulated promoter activity of HHV-8 Rta promoter in BC-3 and HUVEC cells. However, HIV-1 Tat and gp120 failed to act synergistically with interfe ron-γ to enhance Rta promoter activity in HUVECs. Conclusion Cytokine IFN-γ similar to that produced by HIV-1-infected T cells indu ced lytic cycle replication of HHV-8, at less in part, through HHV-8 Rta promo ter.更多还原