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。前期工作 已经证明了I型干扰素(干扰素)诱导的蛋白质限制了灵长类慢病毒在自然和 非自然宿主。事实上，我们之前的研究已经发现了抗病毒干扰素刺激 基因(ISGs)，阐明了它们的作用机制，并揭示了灵长类慢病毒的进化方式 逃避或中和抗病毒蛋白。此外，我们还利用这些知识来产生小说 更好地代表在人类中传播的用于非人类灵长类动物的HIV-1毒株的嵌合病毒 模型，包括一种最低限度修改的HIV-1毒株(StHIV)，当出现以下情况时，可能会导致尾辫猕猴感染艾滋病 在急性感染期间，CD8细胞会瞬间耗尽。在这项新的提案中，我们将探讨 I型干扰素和新型抗病毒ISGs在体外和体内限制灵长类慢病毒复制的作用。具体目标1 将探索抗病毒ISGs影响病毒进入的作用机制。使用CRISPR屏幕，我们有 发现了新的ISGs，其表达似乎抑制了HIV-1感染，特别是在病毒进入步骤。 其中一个基因的敲除会增强HIV-1的感染，其方式根据不同的 产生包膜蛋白的特定菌株。重要的是，ISG对HIV-1的影响程度 特定环境变异体介导的感染与携带该环境变异体的病毒的敏感性相关 对I型干扰素的抑制作用。我们将通过以下方式确定干扰素/ISGs如何抑制HIV-1进入的分子细节 跨细胞类型和物种阐明病毒和宿主抑制的决定因素，并使用成像和其他 确定抑制如何影响传入病毒命运的方法。我们还将确定新奇的ISG 有助于区分原发传播的创始人和慢性HIV-1毒株对干扰素的敏感性 改装/未改装的钻头。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 猕猴体内的I型干扰素以确定干扰素对灵长类慢病毒复制的影响。特别是，我们 将确定I型干扰素阻断对猪尾猕猴stHIV复制和临床病程的影响。我们 还将使用SIV来确定在体外是否表现出不同的I型干扰素敏感性的HIV-1环境变异体， 在活体内也是如此。最后，我们将确定I型干扰素如何影响新城疫病毒的传播和竞争力。 使用条形码SHIV，携带目标1中定义的对干扰素/ISG敏感和耐药的HIV-1环境蛋白，通过 确定在存在和不存在I型干扰素的情况下，每个SIV在猕猴体内的传播效率 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.