INFECTIVITY OF HSIV-VIF CHIMERA IN NEWBORN PIGTAILED MACAQUES

HSIV-VIF 嵌合体在新生短尾猴中的感染性

• 批准号: 8357619
• 负责人: Shiu-Lok Hu
• 金额: $ 37.79万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-05-01 至 2012-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8357619
• 关键词: Acute; Adolescent; Animal Model; Animals; Blood Cells; Chimera organism; Comparative Study; Disease; Engineering; Family suidae; Funding; Grant; HIV-1; Human; Infection; Integration Host Factors; Macaca; Macaca mulatta; Macaca nemestrina; Mediating; Medicine; National Center for Research Resources; Newborn Animals; Newborn Infant; Phase; Pilot Projects; Plasma; Primates; Principal Investigator; Protein Isoforms; Proteins; Research; Research Infrastructure; Resistance; Resources; SIV; Source; Tail; Testing; United States National Institutes of Health; Viral; Viral Load result; Viremia; Virion; Virus; college; cost; improved; in vivo; particle; prevent; vif Genes

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.

该副本是利用资源的众多研究子项目之一 由NIH/NCRR资助的中心赠款提供。对该子弹的主要支持 而且，副投影的主要研究员可能是其他来源提供的 包括其他NIH来源。 列出的总费用可能 代表subproject使用的中心基础架构的估计量， NCRR赠款不直接向子弹或副本人员提供的直接资金。 当前，没有用于HIV-1感染和疾病的动物模型，因为该病毒对人类高度特异。猕猴通常对HIV-1具有抗性，但除猪尾猕猴例外。我们的实验室表明，猪尾猕猴无法表达TRIM5-ALPHA的功能同工型，该功能是限制恒河猴中HIV-1复制的宿主因子。因此，HIV-1可能只需要克服另一个宿主因子APOBEC3G/F即可在猪尾猕猴中成功复制。为了检验这一假设，我们与贝勒医学院的J. Kimata博士合作，该学院设计了一个HIV-1克隆，其中包括Sivmne的VIF基因，使其可以抵消APOBEC3G/F介导的限制。该嵌合病毒HSIV-VIF为96％HIV-1和4％SIV。它像sivmne一样有效地在刺激的猪尾猕猴血细胞中复制。在另一项试点研究中，我们用HSIV-VIF静脉内接种了2个少年猪尾猕猴。尽管两种动物都被感染，但血浆病毒血症在感染后10个月内并未维持。由于新生动物更容易受到慢病毒感染和疾病的影响，因此我们用HSIV-VIF接种了两只新生的猪尾猕猴。两种动物都被感染，峰血浆病毒负荷为0.5-1x105拷贝/ml。尽管急性期感染后病毒载荷降低到基线（100份/mL），但在第28周到44周之间，这两种动物都反弹，表明持续感染。比较研究使我们能够鉴定出Pt-Tropic HIV-1和SIVMNE（1）SIV VIF之间的两个显着差异与PT-热带HIV-1病毒颗粒没有关联。 （2）虽然PT-Tropic HIV-1降解PT APOBEC3G和APOBEC3F，但它比致病性SIVMNE更能防止其纳入病毒体。因此，尽管SIV VIF对于通过PT-Tropic HIV-1持续感染是必需的，但在体内可能需要改善APOBEC3蛋白的表达和抑制APOBEC3蛋白。