Novel interferon dependent innate antiviral mechanisms

新型干扰素依赖性先天抗病毒机制

• 批准号: 8446488
• 负责人: HERBERT W VIRGIN
• 金额: $ 33.23万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至 2015-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8446488
• 关键词: Antiviral Agents; Autophagocytosis; Biochemistry; Crimean-Congo Hemorrhagic Fever Virus; Development; Genes; Goals; Growth; Host resistance; Immune; In Vitro; Infection Control; Interferon-alpha; Interferons; Listeria monocytogenes; Mediating; Mus; Natural Immunity; Norovirus; Process; Proteins; Role; Signal Transduction; Toxic effect; Toxoplasma gondii; Ubiquitin; Viral; Virus; biodefense; in vivo; macrophage; novel; pathogen; programs

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.

这是先前MRCE计划的RP6的延续，现在以PPG的RP5（RP4-RP8）呈现为 分析负责优先级干扰素（IFN）广泛频谱活性的机制 病原体。虽然对IFN信号的了解众所周知，但对IFN诱导的效应子的了解较少 控制感染的机制。我们推测特定IFN效应器的直接操纵 机制可以提供广泛的保护，而无需与IFN给药相关的毒性。我们的目标 是两个方面的：（i）发现新型IFN诱导的抗病毒分子，以及（ii）定义其重要性， 生物化学和机制。我们确定ISG15是IFN诱导的抗病毒分子，证明它是广泛的 抗病毒作用，鉴定出宿主蛋白和ISG15残基对其抗病毒功能必不可少，并鉴定出A 新的新型免疫逃避策略由克里米亚刚果出血病毒在内的多种病毒使用 对抗ISG15和泛素依赖性的先天免疫。在分析ISG15时，我们继续 发现过程，识别自噬基因ATG5对于IFNY诱导的感染的控制至关重要 与单核细胞增生李斯特菌（LM），弓形虫弓形虫（TG）和鼠Norovirus（MNV）。我们建议 通过以下目的继续这两种两条统计的机制和发现方法： AIM 1：IFN诱导的效应分子机制：专注于自噬。 1A）评估ATG5和ATG7在控制感染中使用优先病原体的重要性的一般性。 1b）确定自噬蛋白ATG5和ATG7是否需要IFNA介导的抑制 病毒生长。 1C）定义负责ATG5/7/自噬在IFN诱导的控制中的重要性的机制 病原体复制。 1d）确定IFNY调节自噬的信号传导机制。 AIM 2：IFN诱导的效应分子鉴定。 2a）定义了原发性巨噬细胞中IFNA和IFNY的转录特征。 2B）筛选候选IFN诱导的效应分子用于体内和体外的抗病毒活性。 2C）生成和分析缺乏候选抗病毒分子的小鼠。