CAJ | 학술논문

目的利用腺病毒系统研究HIV-1 Vpr蛋白在T细胞来源的C8166细胞内的高表达以及细胞内该蛋白对T细胞毒性.方法将表达目的蛋白Vpr的重组腺病毒rAd-vpr和空白载体病毒rAd-vector分别感染对数生长期的C8166细胞,流式细胞术(FCM)检测细胞周期分布、凋亡和坏死.用Hoechst-PI荧光染色观察细胞的凋亡和坏死,JC-1荧光染色测定线粒体膜电势.结果 Annexin V-PI染色及Hoechst-PI染色一致显示HIV-1 Vpr能显著诱导C8166细胞凋亡和坏死;PI细胞周期结果表明HIV-1 Vpr能阻滞C8166细胞于G2期;JC-1荧光染色法测定HIV-1 Vpr致C8166线粒体膜电势下降.结论细胞内HIV-1 Vpr介导的T细胞毒性包括线粒体功能障碍、细胞周期的G2期阻滞和细胞的死亡.
목적 이용선병독계통연구HIV-1 Vpr단백재T세포래원적C8166세포내적고표체이급세포내해단백대T세포독성.방법 장표체목적 단백Vpr적중조선병독rAd-vpr화공백재체병독rAd-vector분별감염대수생장기적C8166세포,류식세포술(FCM)검측세포주기분포、조망화배사.용Hoechst-PI형광염색관찰세포적조망화배사,JC-1형광염색측정선립체막전세.결과 Annexin V-PI염색급Hoechst-PI염색일치현시HIV-1 Vpr능현저유도C8166세포조망화배사;PI세포주기결과표명HIV-1 Vpr능조체C8166세포우G2기;JC-1형광염색법측정HIV-1 Vpr치C8166선립체막전세하강.결론 세포내HIV-1 Vpr개도적T세포독성포괄선립체공능장애、세포주기적G2기조체화세포적사망.
Objective To investigate the T cell cytotoxicity induced by recombinant adenovirus carrying HIV-1 vpr gene.Methods C8166 cells infected with rAd-vpr or negative control rAd-vector,were analyzed for cell cycle distribution and cell death by flow cytometry.The discrimination of living cells,apoptotic and necrotic cells were differentiated with Hoechst-PI double staining under the confocal microscopy.Changes of mitochondrial membrane potential(△ψm)were monitored by JC-1 staining method.Results Annexin V-PI and Hoechst-PI staining indicated the death effects of HIV-1 Vpr on C8166 cells.PI flow cytometric analysis showed that cell cycle arrested in G2 phase.C8166 cell△ψm collapse mediated by Vpr was detected by JC-1 fluorescent staining.Conclusion The ability of recombinant adenovirus carrying HIV-1 vpr gene to induce mitochondria dysfunction,cell cycle G2 phase arrest and cell death was confirmed in C8166 cells.