Generation and characterization of new CCR5-tropic HIV-1rmt clones
Generation and characterization of new CCR5-tropic HIV-1rmt clones
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DOI：
10.2152/jmi.64.272
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发表时间：
2017-08-01
影响因子：
0.7
通讯作者：
Nomaguchi, Masako
中科院分区：
文献类型：
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作者：
Doi, Naoya;Sakai, Yosuke;Nomaguchi, Masako
为了开发有效的非人类灵长类动物模型来应对大量的HIV-1/AIDS研究，需要具有各种生物学特性的恒河猴嗜性HIV-1(HIV-1rmt)克隆。这样的克隆，如果有的话，是在实验上阐明HIV-1在宿主个体中的复制和致病性的强大工具，也是开发抗HIV-1药物/疫苗的有力工具。然而，目前只报告了数量有限的HIV-1rmt克隆。在本研究中，我们通过标准的重组DNA和细胞内同源重组技术，获得了携带多种CCR5嗜性env(包膜)基因的新的HIV-1rmt克隆。由此产生的病毒克隆包含来自可诱发艾滋病的实验室或两个临床分离的病毒株的env序列。我们进一步构建了他们在环境中含有N160K、S304G或G310R突变的变异克隆，这些变异克隆可能会改变病毒以使其更好地生长。对新产生的克隆进行了病毒学特性分析，如表达环境病毒、单周期感染性和多周期复制能力。在检查的许多新克隆中，有两个被发现在猕猴细胞中比之前用于比较的构建的克隆生长得更好。我们在这里描述的研究构成了获得CCR5嗜性HIV-1rmt克隆的第一步和必要的一步，这些克隆可用于针对感染者的各种基础和临床研究项目。
To develop effective non-human primate models for coping with numerous HIV-1/AIDS studies, rhesus macaque-tropic HIV-1 (HIV-1rmt) clones with a variety of biological properties are required. Such clones, if available, are powerful tools to experimentally elucidate HIV-1 replication and pathogenicity in host individuals, and also to develop anti-HIV-1 drugs/vaccines. However, only limited numbers of HIV-1rmt clones have been currently reported. In the present study, we generated new HIV-1rmt clones carrying various CCR5-tropic env (envelope) genes by standard recombinant DNA and intracellular homologous recombination techniques. Resultant virus clones contain the env sequences derived from an AIDS-inducible laboratory or two clinically isolated viral strains. We further constructed their variant clones bearing N160K, S304G, or G310R mutation in Env that potentially can change the viruses to better grow. Newly generated clones were analyzed for their virological properties such as Env expression, single-cycle infectivity, and multi-cycle replication ability. Out of a number of new clones examined, two were found to grow better in macaque cells than the previously constructed clone used for comparison. Our study described here constitutes the initial and essential step towards obtaining CCR5-tropic HIV-1rmt clones useful for various basic and clinical research projects on infected individuals.