Survival of influenza A virus infected cells and effects on pathogenesis

 描述(申请人提供)：我们对甲型流感病毒引起疾病的机制非常感兴趣。我们开发了一种新的体内报告系统，用来标记被感染的细胞。重要的是，即使病毒最终从受感染的细胞中清除，这些细胞也会继续表达报告。使用这个系统，我们有 确定了一种肺上皮细胞类型(俱乐部细胞)，可以被感染，然后清除病毒感染并存活下来。在我们的初步数据中，我们已经表明这些细胞对干扰素刺激高度敏感。存活细胞也高度上调趋化因子的表达，这些存活细胞的特异性耗尽或移除对病毒诱导的损伤后的肺修复有积极的影响。这项提议将测试两个主要问题：1)为什么俱乐部细胞能够唯一地直接病毒感染(目标1)和2)存活细胞如何影响病毒感染期间的肺部病理(目标2)。目的1通过描述病毒刺激过程中干扰素刺激基因(ISG)反应的性质，详细介绍旨在了解Club细胞如何在感染中存活的实验。我们不仅将研究影响ISG反应增加的转录和表观遗传因素，还将确定哪些ISG对影响细胞生存最重要。目的2建议研究存活细胞如何在病毒发病机制中发挥作用。我们将对体内存活的细胞进行基因操作，以调节它们分泌免疫调节因子的能力。我们还将直接中和幸存的俱乐部细胞分泌的因子。这是第一次描述可以在急性流感病毒感染中存活的细胞，这项提议中的实验将增加我们的 了解细胞存活的潜在机制以及这些细胞如何在病毒发病中起作用。这些问题不仅是理解病毒如何诱发疾病的基础科学的重要问题，而且也可能为针对存活细胞群体的新型治疗干预策略提供基础。

 DESCRIPTION (provided by applicant): We are broadly interested in the mechanisms of influenza A virus induced disease. We have developed a novel in vivo reporter system with which to label cells that become infected. Importantly, these cells continue to express the reporter even if the virus is eventually cleared from the infected cell. Using this system, we have identified a lung epithelial cell type (club cells) that can become infected, and then clear and survive viral infection. In our preliminary data, we have shown that these cells are highly sensitive to interferon stimulation. Surviving cells also have highly up-regulated expression of chemokines, and that specific depletion or removal of these surviving cells positively influences lung repair after virally induced injury. This proposal will test two major questions: 1) Why are club cells uniquely able to survive direct viral infection (Aim 1) and 2) How are surviving cells influencing lung pathology during viral infection (Aim 2). Aim 1 details experiments designed to understand how club cells are surviving infection by characterizing the nature of the interferon stimulated gene (ISG) response during viral stimulation. We will not only look at the transcriptional and epigenetic factors influencing the increased ISG response, but also define which ISGs are the most important for influencing cellular survival. Aim 2 proposes to study how surviving cells are contributing to viral pathogenesis. We will genetically manipulate surviving cells in vivo to modulate their ability to secrete immunomodulatory factors. We will also directly neutralize the factors secreted by surviving club cells. This is the first description of cells tha can survive acute influenza virus infection, and the experiments in this proposal will increase our understanding of the mechanisms underlying cell survival as well as how these cells contribute to viral pathogenesis. Not only are these important questions for understanding the basic science of how viruses induce disease, but may also provide the basis of novel therapeutic intervention strategies targeting surviving cell populations.