The role of tuft cells in norovirus pathogenesis

簇细胞在诺如病毒发病机制中的作用

基本信息

批准号：
10435511
负责人：
Craig Brian Wilen
金额：
$ 63.81万
依托单位：
YALE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-07-08 至 2025-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10435511
关键词：
Acute Adaptive Immune System Adoptive Transfer Adult Animals Antigen Presentation Antiviral Agents Bacteria Binding Biological Models CD8-Positive T-Lymphocytes Cell Culture System Cell Culture Techniques Cell physiology Cells Cellular Immunity Cessation of life Child Chronic Coculture Techniques Colitis Complex Data Detection Disease Enteral Epithelial Cells Food Gastroenteritis Gastrointestinal tract structure Genes Genetic Goals Helminths Human Immune Immune Evasion Immune response Immune system Immunity Immunologics Impairment In Vitro Infection Inflammatory Bowel Diseases Interferons Intestines Knock-in Mouse Knockout Mice Link Major Histocompatibility Complex Malignant Neoplasms Mediating Modeling Mucosal Immune System Mucosal Immunity Mucous Membrane Mus Nematoda Neurons Norovirus Organoids Pathogenesis Pathologic Population Predisposition Property Recovery Resistance Role Signal Transduction T-Lymphocyte Testing Time Tropism Vaccines Viral Viral Gastroenteritis Virus Virus Diseases Work allergic response antigen detection cell killing cell type chronic infection conditional knockout enteric virus infection gut inflammation gut microbiota immune function in vivo insight microbiome nerve supply novel prevent receptor single-cell RNA sequencing tool transcriptome transmission process vaccine development virus genetics wound

项目摘要

Project Summary Human norovirus is the leading cause of acute gastroenteritis globally causing up to 200,000 deaths per year. However, there are no antiviral drugs or vaccines. We use the closely related mouse or murine norovirus (MNV) as a model system to understand how human norovirus causes infection and disease. More broadly, we also use MNV as a tool to uncover novel and fundamental aspects of how viruses interact with the host immune system and other intestinal microbes including bacteria and worms. We recently discovered CD300lf as a receptor of MNV. We leveraged this finding to identify tuft cells as a target cell of MNV in the mouse intestines. Tuft cells are rare epithelial cells that sense and initiate an immune response against intestinal worms. Tuft cells are also important in regulating intestinal inflammation, wound recovery, and several mucosal cancers. In preliminary data, we developed novel tuft cell deficient mice and both CD300lf conditional knockout and knock-in mice to study the role of tuft cells during acute and chronic norovirus infection. W We also developed an in vitro MNV tuft cell culture system that will enable us to dissect the complex interactions between norovirus, tuft cells, and the immune system. The objectives of this proposal are to determine the role of tuft cells in norovirus infection, to determine the immunological consequences of tuft cell infection, and to discover how infected tuft cells evade CD8+ T cell killing. In Aim 1, we will determine if viral infection impairs tuft cell function. We will identify the role of tuft cells in acute and persistent MNV infection. We will determine how MNV kills tuft cells during chronic infection, how infection perturbs the tuft cell transcriptome, and whether virus infection of tuft cells impairs the ability of tuft cells to protect the host against worm infection. In Aim 2, we will determine the mechanism by which tuft cells act as an immunoprivileged niche for MNV. MNV-specific CD8+ T cells ignore infected tuft cells during chronic infection yet they remain functional and able to detect antigen ex vivo. We will identify how tuft cell tropism enables resistance to CD8+ T cells by combining novel mouse lines, viral genetics, tuft cell and T cell co-culture, and adoptive transfer of both tuft cell and MNV-specific CD8+ T cells. In particular, we will determine if quiescent long-lived tuft cells, a tuft cell subtype, are a novel immunoprivileged reservoir for chronic infection. We anticipate this work will generate a detailed understanding about the cell tropism regulating norovirus transmission, the role of viral infection in type II immunity, and the mechanism by which norovirus evades immune CD8+ T cells. This is critical to our long-term goal of deciphering mechanisms of human norovirus pathogenesis and developing a successful human norovirus vaccine.

项目概要人类诺如病毒是全球急性胃肠炎的主要原因，每年导致多达 20 万人死亡年。然而，没有抗病毒药物或疫苗。我们使用密切相关的小鼠或小鼠诺如病毒（MNV）作为了解人类诺如病毒如何引起感染和疾病的模型系统。更广泛地说，我们还使用 MNV 作为工具来揭示病毒如何与宿主免疫相互作用的新颖和基本方面系统和其他肠道微生物，包括细菌和蠕虫。我们最近发现 CD300lf 作为 MNV 的受体。我们利用这一发现将簇细胞识别为小鼠肠道中 MNV 的靶细胞。簇细胞是罕见的上皮细胞，可以感知并启动免疫对肠道蠕虫的反应。簇细胞在调节肠道炎症、伤口等方面也很重要恢复和几种粘膜癌。在初步数据中，我们开发了新型簇细胞缺陷小鼠和 CD300lf 条件敲除小鼠和敲入小鼠研究簇细胞在急性和慢性诺如病毒感染过程中的作用。我们也开发了一种体外 MNV 簇细胞培养系统，使我们能够剖析之间的复杂相互作用诺如病毒、簇细胞和免疫系统。该提案的目标是确定诺如病毒感染中的簇细胞，以确定簇细胞感染的免疫学后果，以及发现受感染的簇细胞如何逃避 CD8+ T 细胞杀伤。在目标 1 中，我们将确定病毒感染是否会损害簇细胞功能。我们将确定簇绒的作用急性和持续性 MNV 感染中的细胞。我们将确定 MNV 在慢性感染期间如何杀死簇细胞，感染如何扰乱簇细胞转录组，以及簇细胞的病毒感染是否会损害簇细胞的能力细胞保护宿主免受蠕虫感染。在目标 2 中，我们将确定簇细胞作为免疫特权生态位的机制对于MNV。 MNV 特异性 CD8+ T 细胞在慢性感染期间忽略受感染的簇细胞，但它们仍保持功能并能够离体检测抗原。我们将通过以下方式确定簇细胞向性如何能够抵抗 CD8+ T 细胞结合新型小鼠品系、病毒遗传学、簇细胞和 T 细胞共培养以及两种簇细胞的过继转移和 MNV 特异性 CD8+ T 细胞。特别是，我们将确定静态长寿命簇细胞（簇细胞亚型）是否是慢性感染的新型免疫豁免储库。我们预计这项工作将产生对调节诺如病毒的细胞向性的详细了解传播、病毒感染在 II 型免疫中的作用以及诺如病毒逃避免疫的机制 CD8+ T 细胞。这对于我们破译人类诺如病毒机制的长期目标至关重要发病机制和开发成功的人类诺如病毒疫苗。