Autophagy in Antiviral Immunity

抗病毒免疫中的自噬

• 批准号: 7981621
• 负责人: AKIKO IWASAKI
• 金额: $ 39.25万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-05-01 至 2015-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7981621
• 关键词: Ablation; Agonist; Animal Model; Anti-Infective Agents; Antigen Presentation Pathway; Antigens; Antiviral Agents; Autophagocytosis; Autophagosome; Binding; Biochemical; Biological; CD4 Positive T Lymphocytes; Cells; Cytosol; DNA Virus Infections; Data; Dendritic Cells; Endosomes; Family; Fibroblasts; Foundations; Generations; Genes; Genetic; Genetic Transcription; Herpesvirus 1; Homeostasis; Immune response; Immune system; Immunity; In Vitro; Infection; Interferon Type I; Interferon-beta; Interferons; Life; MHC Class II Genes; Measures; Mediating; Mitochondria; Molecular; Mus; Organelles; Pathway interactions; Peptides; Play; Population; Process; Proteins; Proteomics; RNA; RNA Viruses; Reactive Oxygen Species; Receptor Signaling; Reporting; Role; Scientific Advances and Accomplishments; Signal Transduction; Simplexvirus; Starvation; T cell response; TLR7 gene; Techniques; Toll-like receptors; Tretinoin; Vaccine Design; Vesicular stomatitis Indiana virus; Viral; Viral Antigens; Virus; Virus Diseases; Work; adaptive immunity; antigen processing; cell type; combat; cytokine; design; extracellular; human TLR7 protein; immunological intervention; in vivo; melanoma; mouse model; pathogen; promoter; public health relevance; receptor; response; sensor; tool

DESCRIPTION (provided by applicant): Autophagy is an ancient evolutionarily conserved pathway designed to maintain cellular homeostasis by degrading long-lived proteins and organelles in the cytosol. It is also used as a survival mechanism under starvation conditions. Recent studies demonstrated that autophagy is utilized by the cells of the innate and adaptive immune systems to combat viral infections. Innate recognition of viruses occurs via two distinct pathways. In professional viral sensors, the plasmacytoid dendritic cells (pDC), recognition of viruses occurs in the endosomes via Toll-like receptors (TLR) 7 and 9. Our recent work has demonstrated that autophagy plays a key role in recognizing signatures of viral infection in pDCs through TLR7. In contrast to pDCs, most other cell types of the body utilize cytosolic sensors of viral replication via RIG-I like receptor (RLR) family. Molecules involved in autophagy have been shown to block RLR signaling. In addition, recent reports indicate that autophagy delivers endogenous viral antigens to the MHC class II loading compartment, allowing activation of CD4 T cells. However, the relevance of such pathways during in vivo virus infection is unknown. In this application, we present preliminary data that reveal the requirement for Atg5, a key molecule required for formation of autophagosomes, in the transduction of signaling through TLR9 leading to the activation of type I IFN genes in pDCs upon herpes simplex virus (HSV; TLR9 agonist) infection. In addition, we show that autophagy negatively regulates RLR pathway in non-plasmacytoid dendritic cells upon vesicular stomatitis virus infection (VSV; RIG-I agonist). Finally, we demonstrate a key in vivo role for autophagy in the processing and presentation of various forms of antigens on MHC class II in dendritic cells upon HSV-1 infection. Building on these preliminary studies, we propose to examine the importance of autophagy in both innate and adaptive immune responses using a variety of molecular and cell biological techniques and using established mouse models of virus infection. In the first Aim, we will determine the mechanism by which Atg5 and/or autophagy mediates signaling through TLR9 upon HSV infection in pDCs through the use of molecular and cellular biological techniques. In the second Aim, we propose to determine how autophagy regulates RNA sensor activation upon VSV infection through proteomics and biochemical approaches. In the final Aim, we will interrogate how dendritic cells utilize autophagy for processing and presentation of extracellular viral antigens in vitro and in vivo in mice selectively deficient in autophagy within the dendritic cell populations. By providing basic understanding of how autophagy orchestrates the generation of innate and adaptive immunity against virus infections, these studies will help to establish important foundation with which to design vaccines and anti-infective measures against a variety of viral pathogens PUBLIC HEALTH RELEVANCE: While autophagy is an ancient evolutionarily conserved pathway designed to maintain cellular homeostasis by degrading long-lived proteins and organelles in the cytosol, recent studies from our group and others have revealed the role of autophagy in the immune system. In this application, we propose to examine the importance of autophagy in both innate and adaptive antiviral immune responses using well-established genetic, biochemical and cell biological tools as well as in vivo animal models of both RNA and DNA virus infections. The understanding gained from the proposed studies will not only provide scientific advances in how autophagy is utilized by the immune system, but also to help establish critical foundation with which to design immunological interventions and preventative measures against a wide variety of viral diseases.

描述（由申请人提供）：自噬是一种古老的进化保守途径，旨在通过降解细胞质中的长寿命蛋白质和细胞器来维持细胞稳态。它也被用作饥饿条件下的生存机制。最近的研究表明，先天性和适应性免疫系统的细胞利用自噬来对抗病毒感染。病毒的先天识别通过两种不同的途径发生。在专业的病毒传感器——浆细胞样树突状细胞（pDC）中，病毒的识别通过Toll样受体（TLR）7和9发生在核内体中。我们最近的工作表明，自噬在通过TLR7识别pDC中病毒感染特征方面发挥着关键作用。与 pDC 相比，身体的大多数其他细胞类型通过 RIG-I 样受体 (RLR) 家族利用病毒复制的胞质传感器。参与自噬的分子已被证明可以阻断 RLR 信号传导。此外，最近的报告表明，自噬将内源性病毒抗原传递至 MHC II 类装载室，从而激活 CD4 T 细胞。然而，这些途径在体内病毒感染过程中的相关性尚不清楚。在本申请中，我们提供的初步数据揭示了 Atg5 的需求，Atg5 是自噬体形成所需的关键分子，通过 TLR9 进行信号转导，导致单纯疱疹病毒（HSV；TLR9 激动剂）感染后 pDC 中 I 型 IFN 基因的激活。此外，我们还发现，在水泡性口炎病毒（VSV；RIG-I 激动剂）感染后，自噬对非浆细胞样树突状细胞中的 RLR 通路产生负向调节。最后，我们证明了自噬在 HSV-1 感染后树突状细胞中 MHC II 类各种形式抗原的加工和呈递中的关键体内作用。在这些初步研究的基础上，我们建议使用各种分子和细胞生物学技术以及已建立的病毒感染小鼠模型来检查自噬在先天性和适应性免疫反应中的重要性。在第一个目标中，我们将通过使用分子和细胞生物学技术确定 pDC 中 HSV 感染时 Atg5 和/或自噬通过 TLR9 介导信号传导的机制。在第二个目标中，我们建议通过蛋白质组学和生化方法确定自噬如何在 VSV 感染时调节 RNA 传感器激活。在最终目标中，我们将探讨树突状细胞群中选择性缺乏自噬的小鼠体内和体外如何利用自噬来处理和呈递细胞外病毒抗原。通过提供对自噬如何协调针对病毒感染的先天性和适应性免疫的产生的基本了解，这些研究将有助于为设计针对各种病毒病原体的疫苗和抗感染措施奠定重要基础 公共健康相关性：虽然自噬是一种古老的进化保守途径，旨在通过降解细胞质中的长寿命蛋白质和细胞器来维持细胞稳态，但我们小组和其他人最近的研究揭示了自噬在免疫系统中的作用。在此应用中，我们建议使用成熟的遗传、生化和细胞生物学工具以及 RNA 和 DNA 病毒感染的体内动物模型来检查自噬在先天性和适应性抗病毒免疫反应中的重要性。从拟议的研究中获得的理解不仅将为免疫系统如何利用自噬提供科学进展，而且有助于为设计针对多种病毒性疾病的免疫干预和预防措施奠定重要基础。