Autophagy regulation of RSV-induced pulmonary disease

描述(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地识别致病信号并促进适当的非致病反应以清除病毒感染的能力。呼吸道合胞病毒(RSV)是一种特殊的致病病毒，可导致所有年龄段的人发病，特别是在婴儿和有潜在肺部疾病的人中。最近的数据表明，自噬是病原体识别的一个关键的先天免疫途径。这一无处不在的过程为所有细胞提供了不仅保存资源而且将病原体成分运输到适当的免疫识别分子PAMPs的能力。我们的研究已经证实，树突状细胞(DC)对RSV感染的识别依赖于肺内最合适的非致病反应的自噬。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中的关键先天细胞因子来产生有效的免疫反应。我们将使用RSV感染模型的体外和体内分析来检验这一假说，并扩展发现，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，从而有助于反应的严重性。利用特定基因敲除和遗传缺陷的小鼠，包括Beclin+/-和LC3B-/-小鼠，我们将探索自噬机制的缺失如何在体外和体内改变抗RSV反应。随后，我们将通过检测启动自噬小体形成的NAD脱乙酰基酶SIRT1来进一步探索启动自噬反应的重要机制。利用我们建立的RSV感染模型，概括了人类疾病的几个方面，包括粘液高分泌、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，这些研究将探索一些新的和局部的途径，不仅可以进一步确定促进肺粘膜免疫反应的机制，而且可以扩大我们对RSV疫苗设计原理的理解。

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.