Komoto Satoshi, Tsuji Shoutaro, Ibrahim Madiha S, Li Yong-Gang, Warachit Jiranan, Taniguchi Koki, Ikuta Kazuyoshi
Department of Virology, Research Institute for Microbial Diseases, Osaka University, Suita, Osaka 565-0871, Japan.
J Virol. 2003 Oct;77(19):10304-13. doi: 10.1128/jvi.77.19.10304-10313.2003.
Previous data revealed that primary cultures of peripheral blood mononuclear cells (PBMCs) were killed by apoptosis at higher rates after infection with two CRF01_AE primary isolates of human immunodeficiency virus type 1 (HIV-1) than after infection with five other CRF01_AE primary isolates, five subtype B primary isolates, and two subtype B laboratory strains. Here, we show evidence that mutations at the vpu gene which were exclusively identified only in the two CRF01_AE isolates mentioned above are involved in their abilities to induce massive apoptosis in primary CD4(+) T lymphocytes. The rates of virus production by these two isolates in the culture media of infected PBMCs were lower (the same as those of the other CRF01_AE isolates) than those of the subtype B isolates. To confirm the correlation between the higher apoptosis-inducing abilities and the mutations at the vpu gene, infectious molecular clone pNL4-3-based vpu mutants were constructed and examined for their apoptosis induction levels. The apoptosis induction levels after introduction of the vpu mutations were greatly increased in primary CD4(+) T lymphocytes. In contrast, the apoptosis induction abilities of these vpu mutants were lower in human T-cell line MT-4. Thus, the Vpu protein of HIV-1 could play a protective role against virus-induced apoptosis in primary CD4(+) T lymphocytes.
先前的数据显示,与感染另外5株CRF01_AE主要分离株、5株B亚型主要分离株及2株B亚型实验室毒株相比,外周血单个核细胞(PBMC)原代培养物在感染2株1型人类免疫缺陷病毒(HIV-1)CRF01_AE主要分离株后,以更高的速率通过凋亡被杀死。在此,我们证明,仅在上述2株CRF01_AE分离株中发现的vpu基因突变与其在原代CD4(+) T淋巴细胞中诱导大量凋亡的能力有关。这2株分离株在感染的PBMC培养基中的病毒产生率低于B亚型分离株(与其他CRF01_AE分离株相同)。为证实更高的凋亡诱导能力与vpu基因突变之间的相关性,构建了基于感染性分子克隆pNL4-3的vpu突变体,并检测其凋亡诱导水平。在原代CD4(+) T淋巴细胞中,引入vpu突变后的凋亡诱导水平大幅增加。相反,这些vpu突变体在人T细胞系MT-4中的凋亡诱导能力较低。因此,HIV-1的Vpu蛋白可能在原代CD4(+) T淋巴细胞中对病毒诱导的凋亡起到保护作用。