INFECTIVITY OF HSIV-VIF CHIMERA IN NEWBORN PIGTAILED MACAQUES

这个子项目是许多利用资源的研究子项目之一 由NIH/NCRR资助的中心赠款提供。子项目的主要支持 子项目的主要研究者可能是由其他来源提供的， 包括其他NIH来源。 列出的子项目总成本可能 代表子项目使用的中心基础设施的估计数量， 而不是由NCRR赠款提供给子项目或子项目工作人员的直接资金。 目前，还没有HIV-1感染和疾病的动物模型，因为该病毒对人类具有高度特异性。猕猴通常对HIV-1有抵抗力，但猪尾猕猴除外。我们的实验室表明，猪尾猕猴不能表达TRIM 5-alpha的功能亚型，TRIM 5-alpha已被确定为限制HIV-1在恒河猴中复制的宿主因子。因此，HIV-1可能只需要克服另一种宿主因子APOBEC 3G/F的限制，就可以在猪尾猕猴中成功复制。为了验证这一假设，我们与贝勒医学院的J. Kimata博士合作，他设计了一个包含SIVmne的vif基因的HIV-1克隆，使其能够抵消APOBEC 3G/F介导的限制。这种嵌合病毒，HSIV-vif，是96%的HIV-1和4%的SIV。它在刺激的猪尾猕猴血细胞中复制的效率与SIVmne一样。在另一项初步研究中，我们用HSIV-vif静脉内接种2只幼年猪尾猕猴。虽然这两只动物都被感染，但血浆病毒血症在感染后10个月内没有持续。由于新生动物更容易受到慢病毒感染和疾病，我们用HSIV-vif接种了两只新生猪尾猕猴。两只动物均被感染，血浆病毒载量峰值为0.5- 1x 105拷贝/ml。尽管病毒载量在急性期感染后降至基线（100拷贝/ml），但在第28周至第44周期间，两只动物的病毒载量均反弹，表明持续感染。比较研究使我们能够确定Pt-嗜性HIV-1和SIVmne之间的两个显著差异：（1）SIV Vif不与Pt-嗜性HIV-1病毒颗粒相关;（2）虽然Pt-嗜性HIV-1降解Pt APOBEC 3G和APOBEC 3F，但它比致病性SIVmne更小程度地阻止它们包含在病毒体中。因此，虽然SIV Vif对于Pt-嗜性HIV-1的持续感染是必需的，但是APOBEC 3蛋白的改善的表达和抑制对于体内稳健的病毒复制可能是必需的。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Currently, there is no animal model for HIV-1 infection and disease because the virus is highly specific for humans. Macaques are generally resistant to HIV-1, with the exception being pig-tailed macaques. Our lab showed that pig-tailed macaques are unable to express functional isoforms of TRIM5-alpha, which has been identified as a host factor that restricts the replication of HIV-1 in rhesus monkeys. Therefore, it is possible that HIV-1 will only need to overcome restriction by another host factor, APOBEC3G/F, to replicate successfully in pig-tailed macaques. To test this hypothesis, we collaborated with Dr. J. Kimata of Baylor College of Medicine, who engineered an HIV-1 clone that includes the vif gene of SIVmne, allowing it to counteract APOBEC3G/F-mediated restriction. This chimeric virus, HSIV-vif, is 96% HIV-1 and 4% SIV. It replicates in stimulated pig-tailed macaque blood cells as efficiently as SIVmne. In another pilot study, we inoculated 2 juvenile pig-tailed macaques intravenously with HSIV-vif. Although both animals became infected, plasma viremia did not sustain beyond 10 months after infection. Since newborn animals are more susceptible to lentiviral infection and disease, we inoculated two newborn pig-tail macaques with HSIV-vif. Both animals were infected, with a peak plasma viral load of 0.5-1x105 copies/ml. Although viral load decreased to baseline (100 copies/ml) after acute phase infection, it rebounded in both animals between week 28 and 44, indicating persistent infection. Comparative studies allowed us to identified two notable differences between the Pt-tropic HIV-1 and SIVmne (1) SIV Vif does not associate with Pt-tropic HIV-1 viral particles; (2) while Pt-tropic HIV-1 degrades both Pt APOBEC3G and APOBEC3F, it prevents their inclusion in virions to a lesser extent than pathogenic SIVmne. Thus, while SIV Vif is necessary for persistent infection by Pt-tropic HIV-1, improved expression and inhibition of APOBEC3 proteins may be required for robust viral replication in vivo.