Mechanisms of antiviral immunity and tolerance in the intestinal epithelium of Jamaican Fruit Bats

牙买加果蝠肠上皮的抗病毒免疫和耐受机制

• 批准号: 10592671
• 负责人: Diane Bimczok
• 金额: $ 21.46万
• 依托单位: MONTANA STATE UNIVERSITY - BOZEMAN
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-08 至 2025-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10592671
• 关键词: 2019-nCoV; 3-Dimensional; Adult; Antiviral Response; Biological Response Modifiers; Cell Survival; Cell physiology; Cells; Characteristics; Chiroptera; Complex; Coronavirus; Diarrhea; Diet; Disease; Epithelial Cells; Epithelium; Family Pteropodidae; Feces; Filovirus; Fruit; Functional disorder; Gastrointestinal tract structure; Gene Expression; Gene Expression Profile; Gene Expression Profiling; Goals; Henipavirus; Human; Immune system; Immunologics; Immunology; Individual; Infection; Infectious Diseases Research; Influenza; Innate Immune Response; Interferon Activation; Interferon alpha; Interferon-beta; Interferons; Intestines; Jamaican; Kinetics; Maintenance; Mammals; Measures; Middle East Respiratory Syndrome Coronavirus; Modeling; Organ; Organoids; Outcome; Pathogen detection; Pathogenicity; Pathology; Pathway Analysis; Pathway interactions; Pattern; Pattern recognition receptor; Predisposition; Proteome; Proteomics; RNA Viruses; Recombinant Interferon Alfa; Rectum; Resistance; Role; Signal Transduction; Simuliidae; Small Interfering RNA; Small Intestines; Swab; Symptoms; System; Tacaribe virus; Testing; Time; Tissue-Specific Gene Expression; Viral; Virus; Virus Diseases; Virus Replication; Work; Zika Virus; Zoonoses; antiviral immunity; chronic infection; cytotoxicity; enteric pathogen; enteritis; experience; experimental study; gastrointestinal epithelium; genome annotation; human tissue; in vitro Model; influenzavirus; innate immune mechanisms; innovation; innovative technologies; insight; intestinal barrier; intestinal epithelium; model organism; novel; pathogen; pathogen spillover; pathogenic virus; protective pathway; rectal; repair function; response; small molecule inhibitor; spillover event; stem cell derived tissues; transcriptomics

Summary The gastrointestinal (GI) tract is a major target organ for viral infection in bats, but infections rarely cause typical symptoms of viral enteritis such as diarrhea. The long-term goal of our project is to understand the specific innate immune response mechanisms of the intestinal epithelium that enable bats to sustain viral infection without experiencing cytopathic effects and intestinal barrier dysfunction. We hypothesize that protective type I and type III interferon (IFN) responses in Jamaican Fruit Bats (Artibeus jamaicensis, JFBs) enable persistent, asymptomatic viral infection of the gastrointestinal epithelium. To test our hypothesis, we will utilize a novel in vitro model of the JFB intestine, 3-D enteroids, which are complex long-term cultures of primary intestinal epithelial cells generated from adult tissue-derived stem cells. Responses in the gut epithelium of JFBs will be compared to those induced in human enteroids. In Aim 1, we will define type I/III IFN responses of JFB enteroids to viral infection. In Aim 2, we will determine to what extent type I/III IFNs impact viral replication and barrier function in the gastrointestinal epithelium of JFBs. We will analyze intestinal epithelial cell responses to three single-stranded RNA viruses that can infect JFB cells: H18N11 influenza virus, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), and Cedar virus, a non-pathogenic henipavirus. Specifically, we will analyze the kinetics of viral replication and type I/III IFN activation in the enteroids and will also perform a comprehensive proteomics analysis of virus-infected intestinal epithelial cells. To assess the role of virus-induced type I/III IFNs for gastrointestinal epithelial pathology, IFN signaling will be induced or inhibited, and the impact on enteroid susceptibility to viral infection will be analyzed. We also will measure epithelial cell viability, barrier and repair functions that are commonly disrupted during viral enteritis. Our project is technologically innovative, because we will, for the first time, analyze viral infection in enteroid cultures derived from JFBs and because we also will perform a complete proteome analysis of the JFB intestinal epithelium. The proposed work is conceptually innovative, because it will define how virus-induced type I/III IFNs impact intestinal epithelial function and integrity. Our proposed work is significant, because it will provide novel insights in the epithelial cell-intrinsic innate immune mechanisms involved in asymptomatic infection of the bat intestine with viral pathogens, which has important implications for potential pathogen spillover events.

概括 胃肠道（GI）是蝙蝠病毒感染的主要靶器官，但感染很少引起蝙蝠病毒感染。 病毒性肠炎的典型症状如腹泻。我们项目的长期目标是了解 肠上皮的特定先天免疫反应机制使蝙蝠能够维持病毒 感染而不经历细胞病变效应和肠道屏障功能障碍。我们假设 牙买加果蝠 (Artibeus jamaicensis, JFBs) 的保护性 I 型和 III 型干扰素 (IFN) 反应 使胃肠道上皮发生持续的、无症状的病毒感染。为了检验我们的假设，我们将 利用 JFB 肠的新型体外模型，3-D 肠类，它是复杂的长期培养物 由成体组织来源的干细胞产生的原代肠上皮细胞。肠道内的反应 JFB 的上皮细胞将与人类肠样细胞中诱导的上皮细胞进行比较。在目标 1 中，我们将定义 I/III 型 IFN JFB 肠类对病毒感染的反应。在目标 2 中，我们将确定 I/III 型干扰素的影响程度 JFB胃肠道上皮中的病毒复制和屏障功能。我们来分析肠道 上皮细胞对三种可感染 JFB 细胞的单链 RNA 病毒的反应：H18N11 流感 病毒、严重急性呼吸综合征冠状病毒 2 (SARS-CoV-2) 和雪松病毒（一种非致病性病毒） 亨尼帕病毒。具体来说，我们将分析病毒复制和 I/III 型 IFN 激活的动力学 肠类，还将对病毒感染的肠上皮细胞进行全面的蛋白质组学分析。 为了评估病毒诱导的 I/III 型干扰素对胃肠道上皮病理学的作用，干扰素信号传导将被 诱导或抑制，并分析其对肠病毒感染易感性的影响。我们也会 测量病毒性肠炎期间通常被破坏的上皮细胞活力、屏障和修复功能。 我们的项目在技术上是创新的，因为我们将首次分析肠内的病毒感染 来自 JFB 的培养物，因为我们还将对 JFB 进行完整的蛋白质组分析 肠上皮。拟议的工作在概念上是创新的，因为它将定义病毒如何诱导 I/III 型干扰素影响肠上皮功能和完整性。我们提出的工作意义重大，因为它将 为无症状感染所涉及的上皮细胞固有的先天免疫机制提供了新的见解 蝙蝠肠道被病毒病原体感染，这对潜在病原体具有重要意义 溢出事件。