Novel interferon dependent innate antiviral mechanisms

这是先前MRCE计划的RP6的延续，现在作为PPG(Rp4-RP8)的Rp5呈现到 针对优先级分析干扰素(IFN)广谱活性的机制 病原体。虽然对干扰素信号转导知道很多，但对干扰素诱导的效应器知之甚少。 控制感染的机制。我们推测直接操纵特定的干扰素效应器 这些机制可以提供广泛的保护，而不会产生与干扰素注射相关的毒性。我们的目标 是双管齐下的：(I)发现新的干扰素诱导的抗病毒分子，和(Ii)确定它们的重要性， 生物化学和机理。我们鉴定了ISG15是干扰素诱导的抗病毒分子，证明了它的广泛性 抗病毒作用，鉴定了其抗病毒功能所必需的宿主蛋白和ISG15残基，并鉴定了 包括克里米亚刚果出血热病毒在内的几种病毒使用的新免疫逃避策略 以对抗依赖ISG15和泛素的先天免疫。在分析ISG15的同时，我们继续 发现过程，确定自噬基因ATG5是干扰素诱导的感染控制所必需的 与单核细胞增多性李斯特菌(Lm)、弓形虫(Tg)和小鼠诺如病毒(MNV)一起感染。我们建议 通过以下目标继续这两种双管齐下的机制和发现方法： 目的1：干扰素诱导效应的分子机制：关注自噬。 1A)评估ATG5和ATG7在控制优先病原体感染方面的重要性的一般性。 1b)确定IFNA介导的抑制是否需要自噬蛋白ATG5和ATG7 病毒式增长。 1c)确定ATG5/7/自噬在干扰素诱导的控制中的重要性的机制 病原体复制。 1d)确定IFNy调节自噬的信号机制。 目的2：干扰素诱导的效应分子鉴定。 2)确定原代巨噬细胞中IFNA和IFNY的转录特征。 2B)筛选干扰素诱导的体内和体外抗病毒活性的候选效应分子。 2C)产生并分析缺乏候选抗病毒分子的小鼠。

This is a continuation of RP6 of the prior MRCE program, now presented as RP5 of a PPG (RP4-RP8) to analyze the mechanisms responsible for the broad spectrum activities of interferons (IFNs) against priority pathogens. While much is known about IFN signaling, less is known about the IFN-induced effector mechanisms that control infection. We speculate that direct manipulation of specific IFN effector mechanisms may provide broad protection without the toxicity associated with IFN administration. Our goal was two-pronged: (i) to discover novel IFN-induced antiviral molecules, and (ii) to define their importance, biochemistry, and mechanisms. We identified ISG15 as an IFN-induced anti-viral molecule, proved it's broad antiviral role, identified host proteins and ISG15 residues essential for its antiviral function, and identified a novel immune evasion strategy used by several viruses including Crimean Congo Hemorrhagic Fever Virus to counter ISG15-and ubiquitin-dependent innate immunity. While analyzing ISG15, we continued the discovery process, identifying the autophagy gene ATG5 as essential for IFNy-induced control of infection with Listeria monocytogenes (LM), Toxoplasma gondii (TG), and murine norovirus (MNV). We propose to continue this two two-prong approach to mechanism and discovery through the following Aims: Aim 1: IFN-induced effector molecule mechanisms: Focus on autophagy. 1a) Assess the generality of the importance of Atg5 and Atg7 in control of infection with priority pathogens. 1b) Determine whether autophagy proteins Atg5 and Atg7 are required for IFNa-mediated inhibition of viral growth. 1c) Define the mechanisms responsible for the importance of Atg5/7/autophagy in IFN-induced control of pathogen replication. 1d) Identify signaling mechanisms by which IFNy regulates autophagy. Aim 2: IFN-induced effector molecule identification. 2a) Define the transcriptional signature of IFNa and IFNy in primary macrophages. 2b) Screen candidate IFN-induced effector molecules for antiviral activities in vivo and in vitro. 2c) Generate and analyze mice lacking candidate antiviral molecules.