VIF Anatagonists: Antiviral Evaluation in Vitro, Inhibitor resistance and Mole

Vif对于HIV-1和SIV等灵长类慢病毒的体内复制是必不可少的。在过去两年里， 人们已经对Vif支持病毒复制的机制有了相当大的了解。 VIF可抵消胞苷脱氨酶APOBEC 3G和3F的抗病毒作用。为了抑制病毒复制， 这些胞苷脱氨酶必须包装在病毒粒子中，在那里它们编辑新生病毒的负链。 目的基因在靶细胞中的表达。VIF通过促进APOBEC 3G和3F的抗病毒活性而中和其抗病毒活性 蛋白酶体降解到不足以包装成病毒粒子的程度。 该项目的总体目标是确定和开发Vif拮抗剂，可用于评估 Vif拮抗剂的体内抗病毒作用。项目2将优先处理Vif拮抗者 根据抗病毒活性和特异性筛选项目1的抑制剂。因为最有利的VIF 拮抗剂将在SIV致病机制和SIV诱导的脑炎的猕猴模型中进行评估， 抗病毒活性将在与中枢神经系统感染相关的细胞中进行评估，即原代巨噬细胞。 项目2还将评估抑制剂在体外和体内逃逸的机制。项目的主要活动 2包括： 在表达APOBEC(非允许细胞)和APOBEC的细胞中评估抗病毒活性和特异性 不表达APOBEC(允许细胞)的细胞。 中枢神经系统相关靶细胞，即原代巨噬细胞体外抗病毒活性的评价。 通过评估Vif拮抗剂对病毒粒子的影响确定Vif拮抗机制 APOBEC的包装及APOBEC介导的病毒DNA编辑 体内外Vif基因突变对抑制剂的抗性鉴定。 总而言之，这些研究将使我们能够确定最有希望的主要Vif拮抗剂，并将 将在SIV复制和SIV诱导的实验模型中评估的抑制剂的选择 猕猴的脑炎。

Vif is essential to the replication of primate lentiviruses such as HIV-1 and SIV in vivo. In the last two years, a considerable amount of insight has been gained into the mechanism by which Vif supports viral replication. Vif counteracts the antiviral effects of the cytidine deaminases apobec 3G and 3F. To inhibit viral replication, these cytidine deaminases must be packaged within virions where they edit the minus strand of nascent viral cDNA in the target cell. Vif counteracts the antiviral activity of apobec 3G and 3F by promoting their proteasomal degradation to the extent that there are insufficient levels for packaging into virions. The overall goal of this project is to identify and develop Vif antagonists that can be used to assess the antiviral impact of Vif antagonism in vivo. Project 2 will prioritize Vif antagonists that emerge from the inhibitor screen of Project 1 on the basis of antiviral activity and specificity. Since the most favorable Vif antagonists will be evaluated in the macaque model of SIV pathogenesis and SIV-induced encephalitis, antiviral activity will be evaluated in cells that are relevant to CNS infection namely, primary macrophages. Project 2 will also assess mechanisms of inhibitor escape in vitro and in vivo. The major activities of Project 2 include: Evaluation of antiviral activity and specificity in cells that express apobec (non-permissive cells) and cells that do not express apobec (permissive cells). Evaluation of antiviral activity in CNS-relevant target cells in vitro, namely, primary macrophages. Identification of the mechanism of Vif antagonism by assessing impact of Vif antagonists on virion encapsidation of apobec and on apobec-mediated editing of viral cDNA. Identification of mutations in Vif that confer inhibitor resistance in vitro and in vivo. Collectively, these studies will allow us to identify the most promising lead Vif antagonists and prioritize the selection of inhibitors that will be evaluated in an experimental model of SIV replication and SIV-induced encephalitis in macaques.