Effects of Interferon on primate lentiviruses

摘要 该提案是一个多PI提案，旨在了解抗病毒蛋白如何限制宿主范围。先前工作 已经证明I型干扰素（IFN）诱导的蛋白质限制灵长类慢病毒在自然和 非自然宿主事实上，我们以前的研究已经发现了抗病毒干扰素刺激的 基因（ISGs），阐明了它们的作用机制，并揭示了灵长类慢病毒进化的方式 逃避或抵消抗病毒蛋白。此外，我们还利用这些知识来产生新的 更好地代表在人类中传播的HIV-1毒株的嵌合病毒，用于非人灵长类动物 模型，包括一种最小修饰的HIV-1株（stHIV），当 在急性感染期间，CD 8+细胞短暂耗尽。在这个新的建议中，我们将探讨 I型IFN和新型抗病毒ISG在体外和体内限制灵长类慢病毒复制。具体目标1 将探索抗病毒ISG影响病毒进入的作用机制。使用CRISPR筛选， 发现了新的ISG，其表达似乎抑制HIV-1感染，特别是在病毒进入步骤。 敲除这些基因之一会增强HIV-1感染，其方式根据不同的人而有很大不同。 Env蛋白来源的特定菌株。重要的是，ISG对HIV-1的影响程度 由特定Env变体介导的感染与携带该Env变体的病毒的敏感性相关 I型干扰素的抑制作用。我们将确定IFN/ISGs如何抑制HIV-1进入的分子细节， 阐明跨细胞类型和物种的抑制的病毒和宿主决定因素，并使用成像和其他 方法来确定抑制如何影响传入病毒的命运。我们还将确定新的ISG 有助于原发性传播创始者和慢性HIV-1毒株的IFN敏感性差异， 适应性/非适应性SHIV。在具体目标2中，我们将使用新开发的更有效的方法来扰动 I型IFN在猕猴中的作用，以确定IFN对其中灵长类慢病毒复制的影响。我们尤其 将确定I型IFN阻断对猪尾猕猴中stHIV复制和临床过程的影响。我们 还将使用SHIV来确定是否HIV-1 Env变体在体外表现出不同的I型IFN敏感性， 在体内也是如此。最后，我们将确定I型干扰素如何影响SHIV传播和竞争力 使用目标1中定义的带有IFN/ISG敏感性和耐药性HIV-1 Env蛋白的条形码SHIV， 确定在存在和不存在I型IFN的情况下每种SHIV在猕猴中传播的效率 封锁

ABSTRACT This proposal is a multi PI proposal that aims to understand how antiviral proteins limit host range. Prior work has demonstrated that type I interferon (IFN) induced proteins limit primate lentivirus replication in natural and non-natural hosts. Indeed, our previous studies have led to the discovery of antiviral interferon stimulated genes (ISGs), elucidated their mechanism of action and uncovered ways in which primate lentiviruses evolve evade or counteract antiviral proteins. Additionally, we have exploited this knowledge to generate novel chimeric viruses that better represent the HIV-1 strains circulating in humans for use in non-human primate models, including a minimally modified HIV-1 strain (stHIV) that can cause AIDS in pigtail macaques when CD8+ cells are transiently depleted during the acute infection. In this new proposal, we will explore the role of type I IFN and novel antiviral ISGs in limiting primate lentivirus replication in vitro and in vivo. Specific Aim 1 will explore the mechanism of action of antiviral ISGs affecting viral entry. Using a CRISPR screen, we have found novel ISGs whose expression appears to inhibit HIV-1 infection, specifically at the virus entry step. Knockout of one of these genes enhances HIV-1 infection in a manner that varies dramatically according to the particular strain from which the Env protein is derived. Importantly, the magnitude of effect of the ISG on HIV-1 infection mediated by a particular Env variant correlates with the sensitivity of a virus carrying that Env variant to inhibition by type I IFN. We will determine the molecular details of the how IFN/ISGs inhibits HIV-1 entry by elucidating viral and host determinants of inhibition, across cell types and species, and use imaging and other approaches to determine how inhibition affects incoming viron fate. We will also determine how novel ISGs contribute to the differential IFN sensitivity of primary transmitted founder and chronic HIV-1 strains and adapted/unadapted SHIVs. In Specific Aim 2, we will use newly developed, more effective methods to perturb type I IFN in macaques to determine the effect of IFN on primate lentivirus replication therein. In particular, we will determine the effect of type I IFN blockade on stHIV replication and clinical course in pigtail macaques. We will also use SHIV to determine whether HIV-1 Env variants that exhibit differential type I IFN sensitivity in vitro, also do so in vivo. Finally, we will determine how type I IFN affects SHIV dissemination and competitiveness use barcoded SHIVs, bearing IFN/ISG-sensitive and resistant HIV-1 Env proteins, defined in Aim 1, by determining how efficiently each SHIV disseminates in macaques in the presence and absence of type I IFN blockade.