Laboratory of Primate Model, Experimental Research Center for Infectious Diseases, Institute for Virus Research, Kyoto University, Kyoto 606-8507, Japan.
Virology. 2013 Feb 5;436(1):100-11. doi: 10.1016/j.virol.2012.10.036. Epub 2012 Dec 6.
A new simian-human immunodeficiency virus (SHIV), carrying env from an uncloned HIV-1 subtype C clinical isolate (97ZA012), was generated through intracellular homologous recombination, a DNA repair mechanism of the host cell. PCR fragments amplified from an existing SHIV plasmid (a 7-kb fragment from the 5' end and a 1.5-kb fragment from the 3' end) and a 4-kb fragment amplified from 97ZA012 cDNA containing env were co-transfected to human lymphoid cells. The resulting recombinant was subjected to serial passage in rhesus peripheral blood mononuclear cells (RhPBMCs). The resulting SHIV 97ZA012 was replication competent in RhPBMCs and monkey alveolar macrophages, and possessed CCR5 preference as an entry co-receptor. Experimental infection of rhesus macaques with SHIV 97ZA012 caused high titers of plasma viremia and a transient but profound depletion of CD4(+) T lymphocytes in the lung. Animal-to-animal passage was shown to be a promising measure for further adaptation of the virus in monkeys.
一种新的猿猴-人免疫缺陷病毒（SHIV），携带来自未经克隆的 HIV-1 亚型 C 临床分离物（97ZA012）的 env，通过细胞内同源重组产生，这是宿主细胞的一种 DNA 修复机制。从现有的 SHIV 质粒（来自 5'端的 7kb 片段和来自 3'端的 1.5kb 片段）和包含 env 的 97ZA012 cDNA 扩增的 4kb 片段扩增的 PCR 片段被共转染到人类淋巴样细胞中。产生的重组体在恒河猴外周血单核细胞（RhPBMCs）中进行连续传代。产生的 SHIV 97ZA012 在 RhPBMCs 和猴肺泡巨噬细胞中具有复制能力，并具有 CCR5 作为进入共受体的偏好性。用 SHIV 97ZA012 感染恒河猴导致血浆病毒血症的高滴度和肺部 CD4(+) T 淋巴细胞的短暂但严重耗竭。已证明动物间传代是病毒在猴子中进一步适应的一种很有前途的措施。
