Autophagy regulation of RSV-induced pulmonary disease

描述（由申请人提供）：肺中抗病毒免疫反应的调节依赖于有效和适当识别致病信号并促进适当和非致病反应以清除病毒感染的能力。呼吸道合胞病毒（RSV）是一种致病性特别强的病毒，可在所有年龄段引起发病，特别是在婴儿和那些有潜在肺部疾病的人群中。最近的数据表明，自噬是一种关键的先天免疫途径，对病原体识别具有功能。这个无所不在的过程不仅为所有细胞提供了保存资源的能力，而且还将病原体成分运送到适当的免疫识别分子，PAMPs。我们的研究已经确定，树突状细胞（DC）对RSV感染的识别依赖于自噬，这是肺部最合适的非致病性反应。因此，我们的总体假设是RSV感染反应依赖于自噬，通过诱导肺内APC中关键的先天细胞因子产生有效的免疫反应。我们将通过体外和体内分析RSV感染模型来验证这一假设，并扩展研究结果，以证明自噬介导的病毒感染识别决定了获得性免疫反应的方向，从而增加了反应的严重性。利用特异性基因敲除和基因缺陷小鼠，包括Beclin +/-和LC3B-/-小鼠，我们将探索缺乏自噬机制如何改变体外和体内的抗rsv反应。随后，我们将通过检测NAD去乙酰化酶sirt1进一步探索自噬反应启动的重要机制，sirt1已知可启动自噬体的形成。利用我们建立的RSV感染模型概括了人类疾病的几个方面，包括粘液分泌过多、生理变化（AHR）和致病细胞因子谱，这些研究将能够彻底表征与这些关键途径相关的机制变化。因此，这些研究将探索几种新的局部途径，不仅可以进一步确定促进肺粘膜免疫反应的机制，而且可以扩展我们对RSV疫苗设计的基本原理的理解。

DESCRIPTION (provided by applicant): The regulation of anti-viral immune responses in the lung is dependent upon the ability to efficiently and appropriately recognize pathogenic signals and promote the proper and non-pathogenic response to clear the viral infection. Respiratory syncytial virus (RSV) is an especially pathogenic virus that can induce morbidity at all ages, especially in infants and those with underlying lung conditions. Recent data has indicated that a critical innate immune pathway that is functional for pathogen recognition is autophagy. This omnipresent process provides all cells the ability to not only preserve resources but transport pathogen components to the proper immune recognition molecules, PAMPs. Our studies have identified that recognition of RSV infection by dendritic cells (DC) depends upon autophagy for the most appropriate, nonpathogenic response in the lungs. Thus, our overall hypothesis is that RSV infection responses depend upon autophagy to generate an effective immune response through the induction of critical innate cytokines in APC within the lungs. We will test this hypothesis using both in vitro and in vivo analyses with our RSV infection model and extend the findings to demonstrate that autophagy-mediated recognition of the viral infection dictates that direction of the acquired immune response, lending to the severity of the response. Using specific gene knockdown and genetically deficient mice, including Beclin +/- and LC3B-/- mice, we will explore how the absence of autophagy mechanisms alters anti-RSV responses in vitro and in vivo. Subsequently, we will further explore an important mechanism of initiation of the autophagy responses by examining a NAD deacetylase, sirt1, known to initiate autophagosome formation. Using our established model of RSV infection that recapituates several aspects of human disease, including mucus hypersecretion, physiologic changes (AHR) and a pathogenic cytokine profile, these investigations will be able to thoroughly characterize the mechanistic changes associated with these critical pathways. Thus, these studies will explore several novel and topical pathways that can not only further define the mechanisms that promote pulmonary mucosal immune responses, but expand our understanding of rationale vaccine design for RSV.