Generation of Rhesus Macaque-Tropic HIV-1 Clones That Are Resistant to Major Anti-HIV-1 Restriction Factors
Generation of Rhesus Macaque-Tropic HIV-1 Clones That Are Resistant to Major Anti-HIV-1 Restriction Factors
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DOI：
10.1128/jvi.01549-13
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发表时间：
2013-11-01
影响因子：
5.4
通讯作者：
Adachia, Akio
中科院分区：
文献类型：
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作者：
Nomaguchi, Masako;Yokoyama, Masaru;Adachia, Akio
人类免疫缺陷病毒1型（HIV-1）在猕猴细胞中的复制主要受到抗病毒细胞APOBEC3、TRIM5 α /TRIM5CypA和tetherin蛋白的限制。在基础和临床HIV-1/AIDS研究中，我们和其他人已经努力构建嗜猕猴HIV-1 （HIV-1mt）。尽管恒河猴通常被成功地用作感染模型，但迄今为止还没有适合恒河猴体内研究的HIV-1衍生物。在本研究中，为了获得对主要限制性因子具有抗性的新型HIV-1mt克隆，我们改变了先前描述的最佳HIV-1mt克隆的Gag和Vpu。首先，通过序列和结构导向诱变，发现gag -衣壳（CA）中的三个氨基酸残基（M94L/R98S/G114Q）在猕猴细胞系中促进了病毒的生长。携带这些替换的HIV-1mt体外TRIM5 α敏感性测试结果与猕猴外周血单个核细胞（PBMCs）中不同TRIM5等位基因的病毒复制潜力增加密切相关，表明HIV-1mt CA中的三个氨基酸参与了与TRIM5 α的相互作用。其次，我们将该克隆Vpu的跨膜结构域替换为猴免疫缺陷病毒SIVgsn166 Vpu的相应区域。得到的克隆MN4/LSDQgtu能够拮抗猕猴tetherin，但不能拮抗人tetherin，其Vpu在猕猴细胞系中有效地发挥了病毒复制的作用。值得注意的是，MN4/LSDQgtu在恒河猴PBMCs中的生长与SIVmac239相当，并且比我们的任何其他HIV-1mt克隆都要好得多。总之，MN4/LSDQgtu是第一个在恒河猴细胞中表现出对主要限制性因子抵抗的HIV-1衍生物。
Human immunodeficiency virus type 1 (HIV-1) replication in macaque cells is restricted mainly by antiviral cellular APOBEC3, TRIM5 alpha/TRIM5CypA, and tetherin proteins. For basic and clinical HIV-1/AIDS studies, efforts to construct macaque-tropic HIV-1 (HIV-1mt) have been made by us and others. Although rhesus macaques are commonly and successfully used as infection models, no HIV-1 derivatives suitable for in vivo rhesus research are available to date. In this study, to obtain novel HIV-1mt clones that are resistant to major restriction factors, we altered Gag and Vpu of our best HIV-1mt clone described previously. First, by sequence-and structure-guided mutagenesis, three amino acid residues in Gag-capsid (CA) (M94L/R98S/G114Q) were found to be responsible for viral growth enhancement in a macaque cell line. Results of in vitro TRIM5 alpha susceptibility testing of HIV-1mt carrying these substitutions correlated well with the increased viral replication potential in macaque peripheral blood mononuclear cells (PBMCs) with different TRIM5 alleles, suggesting that the three amino acids in HIV-1mt CA are involved in the interaction with TRIM5 alpha. Second, we replaced the transmembrane domain of Vpu of this clone with the corresponding region of simian immunodeficiency virus SIVgsn166 Vpu. The resultant clone, MN4/LSDQgtu, was able to antagonize macaque but not human tetherin, and its Vpu effectively functioned during viral replication in a macaque cell line. Notably, MN4/LSDQgtu grew comparably to SIVmac239 and much better than any of our other HIV-1mt clones in rhesus macaque PBMCs. In sum, MN4/LSDQgtu is the first HIV-1 derivative that exhibits resistance to the major restriction factors in rhesus macaque cells.