Regulation of antiviral immunity at barrier surfaces by interleukin-1 family cytokines

项目摘要 我的长期目标是在一个认可的学术机构获得终身职位， 在病毒学和先天免疫学之间的界面进行独立研究。我设想发展 一项研究计划，利用新的方法来研究屏障抗病毒免疫的机制 在一些实施方案中，所述方法包括在皮肤、肠、肺和口腔粘膜的表面上施用抗生素。获得独立的NIH途径 K99/R 00奖将为我提供额外的培训，我需要作为一个独立的调查成功。在 在这个建议中，我概述了一个培训计划，包括正式的课程，参与职业生涯， 发展课程和讲习班，以及与咨询委员会成员的科学合作。的技能 我将在该奖项的指导阶段获得将使我唯一有资格回答重要的 与宿主对病毒感染的反应有关的问题。 这项研究的科学目标是了解非造血细胞如何有助于抗病毒 屏障表面的抗扰度。屏障表面是各种病毒进入的主要途径， 在这些部位引发的抗病毒应答可对病毒在受感染宿主内的传播具有显著影响。 许多感染屏障表面的病毒最初在上皮细胞内复制。然而，抗病毒药物 由这些细胞诱导的反应及其在防止病毒从感染的原发部位扩散中的作用是 不太了解。上皮细胞中抗病毒免疫的研究主要集中在以下方面的作用： 干扰素（IFN）。然而，许多病毒编码阻断IFN表达的蛋白质。因此，我建议细胞 有替代机制，以沟通感染和限制病毒复制，在遇到 阻断IFN表达的病毒。这个建议是基于我的观察， 表达促进白细胞介素（IL）-1细胞因子从感染的上皮细胞释放，并且IL-1 细胞因子在人成纤维细胞中引发抗病毒状态。我假设上皮细胞感染了干扰素- 逃避病毒释放预先形成的IL-1细胞因子，以诱导邻近的非- 造血细胞此外，我认为屏障组织中的局部IL-1 R信号在 控制病毒从最初感染部位的传播。在本提案中，我将探讨该模型的关键方面 抗病毒免疫力！具体来说，我将确定IL-1机制的遗传和生化基础， 从病毒感染的上皮细胞释放细胞因子（目的1），确定的机制，其中一个关键的 IL-1细胞因子诱导的抗病毒应答的调节因子，转录因子IRF 1，被激活或抑制 (Aim 2），并定义非造血细胞IL-1受体（IL-1 R）信号传导在抗病毒治疗中的抗病毒机制。 人皮肤（目标3）。这些研究的结果将提供重要的洞察力， 宿主和病毒之间的关系，并将最终为我作为一个独立的成功职业生涯奠定基础。 调查员

Project Summary My long-term goal is to obtain a tenure track position at an accredited academic institution where I will perform independent research at the interface between virology and innate immunology. I envision developing a research program that utilizes novel approaches to investigate mechanisms of antiviral immunity at barrier surfaces, such as those of the skin, gut, lung, and oral mucosa. Obtaining the NIH pathway to independence K99/R00 award will provide me with the additional training I need to succeed as an independent investigator. In this proposal, I outline a training plan that encompasses formal coursework, participation in career development courses and workshops, and scientific collaboration with advisory committee members. The skills that I will acquire during the mentored phase of the award will make me uniquely qualified to answer important questions related to the host response to viral infection. The scientific goal of this study is to understand how non-hematopoietic cells contribute to antiviral immunity at barrier surfaces. Barrier surfaces are primary routes of entry for diverse viruses, and the initial antiviral responses elicited at these sites can have marked effects on viral spread within an infected host. Many viruses that infect barrier surfaces initially replicate within epithelial cells. However, the antiviral responses induced by these cells and their roles in preventing viral spread from the primary site of infection are poorly understood. Studies of antiviral immunity in epithelial cells have predominately focused on the role of interferons (IFNs). Yet, many viruses encode proteins that block IFN expression. Therefore, I propose that cells have alternative mechanisms to communicate infection and restrict virus replication during encounters with viruses that block IFN expression. This proposal is founded on my observation that viruses that inhibit IFN expression promote the release of interleukin (IL)-1 cytokines from infected epithelial cells, and that IL-1 cytokines elicit an antiviral state in human fibroblasts. I hypothesize that epithelial cells infected with IFN- evading viruses release preformed IL-1 cytokines to induce an antiviral response in neighboring non- hematopoietic cells. In addition, I propose that local IL-1R signaling in barrier tissues plays an important role in controlling virus spread from the initial site of infection. In this proposal, I will explore key aspects of this model of antiviral immunity.! Specifically, I will determine the genetic and biochemical basis for the mechanism of IL-1 cytokine release from virus-infected epithelial cells (Aim 1), determine the mechanisms by which a critical regulator of IL-1 cytokine induced antiviral responses, the transcription factor IRF1, is activated or inhibited (Aim 2), and define the antiviral mechanism(s) of non-hematopoietic cell IL-1 receptor (IL-1R) signaling in human skin (Aim 3). The results of these studies will provide important insight into the earliest encounters between host and virus, and will ultimately set the framework for my successful career as an independent investigator.