Genetic Identity and Biological Phenotype of a Transmitted/Founder Virus Representative of Nonpathogenic Simian Immunodeficiency Virus Infection in African Green Monkeys
Genetic Identity and Biological Phenotype of a Transmitted/Founder Virus Representative of Nonpathogenic Simian Immunodeficiency Virus Infection in African Green Monkeys
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DOI：
10.1128/jvi.01603-10
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发表时间：
2010-12-01
影响因子：
5.4
通讯作者：
Kirchhoff, Frank
中科院分区：
文献类型：
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作者：
Gnanadurai, Clement Wesley;Pandrea, Ivona;Kirchhoff, Frank
了解非致病性猴免疫缺陷病毒（SIV）感染中疾病进展的缺乏对于解读人类艾滋病的免疫发病机制至关重要。然而，在体内的研究受到了阻碍，缺乏感染性分子克隆（IMC）的SIV适合解剖的病毒和宿主因素负责的非致病性表型。在这里，我们描述的识别，克隆和生物学分析的第一个传播/创始人（T/F）病毒代表一个非致病性SIV感染。在病毒血症峰值时，从静脉接种从未在体外繁殖的非洲绿色猴SIV（SIVagm）毒株（Sab 92018）的急性感染的萨比乌斯猴（绿猴）中采集血液。为了产生IMC，我们首先使用常规（批量）PCR从外周血单核细胞（PBMC）DNA扩增全长病毒基因组。虽然这产生了两个完整的SIVagmSab基因组，但生物学表征显示两者都是复制缺陷型的。然后，我们进行单基因组扩增（SGA）以从血浆病毒RNA产生部分重叠的5'（n = 10）和3'（n = 13）半基因组。这些扩增子的分析揭示了代表T/F病毒后代的几乎相同的病毒序列簇。合成其中之一的共有序列产生IMC（Sab 92018 ivTF），其产生感染性CCR 5嗜性病毒体，并在Molt-4克隆8细胞和非洲绿色猴PBMC中复制至高滴度。Sab 92018 ivTF还在萨白对虾猴中引发了生产性感染，并忠实地再现了亲本Sab 92018菌株的复制动力学和非致病性表型。因此，这些结果将T/F病毒的概念扩展到非致病性SIV感染，并提供了一个重要的新工具来定义疾病非进展的病毒决定因素。
Understanding the lack of disease progression in nonpathogenic simian immunodeficiency virus (SIV) infections is essential for deciphering the immunopathogenesis of human AIDS. Yet, in vivo studies have been hampered by a paucity of infectious molecular clones (IMCs) of SIV suitable to dissect the viral and host factors responsible for the nonpathogenic phenotype. Here, we describe the identification, cloning, and biological analysis of the first transmitted/founder (T/F) virus representing a nonpathogenic SIV infection. Blood was collected at peak viremia from an acutely infected sabaeus monkey (Chlorocebus sabaeus) inoculated intravenously with an African green monkey SIV (SIVagm) strain (Sab92018) that had never been propagated in vitro. To generate IMCs, we first used conventional (bulk) PCR to amplify full-length viral genomes from peripheral blood mononuclear cell (PBMC) DNA. Although this yielded two intact SIVagmSab genomes, biological characterization revealed that both were replication defective. We then performed single-genome amplification (SGA) to generate partially overlapping 5' (n = 10) and 3' (n = 13) half genomes from plasma viral RNA. Analysis of these amplicons revealed clusters of nearly identical viral sequences representing the progeny of T/F viruses. Synthesis of the consensus sequence of one of these generated an IMC (Sab92018ivTF) that produced infectious CCR5-tropic virions and replicated to high titers in Molt-4 clone 8 cells and African green monkey PBMCs. Sab92018ivTF also initiated productive infection in sabaeus monkeys and faithfully recapitulated the replication kinetics and nonpathogenic phenotype of the parental Sab92018 strain. These results thus extend the T/F virus concept to nonpathogenic SIV infections and provide an important new tool to define viral determinants of disease nonprogression.