Rotavirus interaction with gut intraepithelial lymphocytes

轮状病毒与肠上皮内淋巴细胞的相互作用

• 批准号: 10738962
• 负责人: Siyuan Ding
• 金额: $ 27.21万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-01 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10738962
• 关键词: Ablation; Anatomy; Animal Model; Antiviral Agents; Antiviral Response; Aryl Hydrocarbon Receptor; Attenuated; Biological Assay; Biological Models; CD4 Positive T Lymphocytes; CX3CL1 gene; Cell Communication; Cells; Cellular biology; Cessation of life; Child; Coculture Techniques; Country; Cryoultramicrotomy; Data; Development; Diarrhea; Disease; Enteral; Enzyme-Linked Immunosorbent Assay; Epithelial Cells; Epithelium; Etiology; Flow Cytometry; Foundations; Gastrointestinal tract structure; Goals; Host Defense; Hour; Human; Image; Immune Evasion; Immune signaling; Immunity; Immunofluorescence Immunologic; Immunologics; Infant; Infection; Inflammatory; Intestines; Knock-out; Knockout Mice; Knowledge; Life; LoxP-flanked allele; Lymphocyte; Lymphocyte Function; Lymphocyte Subset; Maintenance; Measures; Mediating; Mediator; Methods; Mucous Membrane; Mus; Neonatal; Nonstructural Protein; Organoids; Outcome; Pathologic; Play; Population; Primary Infection; Production; Publications; Recombinants; Resolution; Role; Rotavirus; Rotavirus Infections; Rotavirus Vaccines; Signal Pathway; Small Intestines; System; T-Cell Depletion; T-Lymphocyte Subsets; Testing; Therapeutic; Therapeutic Intervention; Transcript; Vaccine Design; Validation; Viral; Viral Genome; Viral Pathogenesis; Viral Physiology; Virus; Virus Shedding; antagonist; antiviral immunity; chemokine; conditional knockout; design; enteric infection; enteric virus infection; expectation; experimental study; functional outcomes; gut inflammation; improved; in vivo; infancy; innovation; intestinal epithelium; intraepithelial; knockout gene; monolayer; mortality; mouse model; neonatal mice; novel; novel strategies; novel therapeutics; novel vaccines; pathogen; pathogenic virus; prevent; pup; rational design; recruit; response; reverse genetics; single cell mRNA sequencing; single-cell RNA sequencing; vaccine candidate; vaccine efficacy

Project Summary Small bowel intestinal epithelial cells (IECs) are the first line of defense against human enteric viruses, the most common and leading causes of diarrhea and death in infants and young children. How IECs communicate with intraepithelial lymphocytes (IELs) in the small intestine and orchestrate antiviral responses is heavily understudied. Our overall objectives are to better define the host immune signaling pathways in the gastrointestinal tract during infections and to use that information to develop therapeutic interventions to alleviate diarrhea and sequelae. Our preliminary results contrasted our expectation and demonstrated that the numbers of two IEL subsets are significantly reduced during early rotavirus infection in vivo. We also found that rotavirus infection alters the expression of several pro-inflammatory chemokines in infected human and mouse IECs. Based on these data and prior publications, we hypothesize that the IELs are important mediators of host defense against rotavirus infection and that rotavirus-encoded factors antagonize the antiviral activity of IELs in the host small intestine via inhibition of chemokine expression. Testing these hypotheses is currently hampered by the lack of suitable model systems. Accordingly, we have developed a highly tractable murine rotavirus reverse genetics method, a pathologically relevant neonatal mouse model, and several innovative primary human and murine small bowel organoid culturing systems, which will provide an unprecedented resolution of understanding of IEC-IEL crosstalk in the context of enteric viral infections. In Aim 1, we will define a functional antiviral role of IELs in rotavirus infection in vivo using immunological approaches and gene knockout mice. In Aim 2, we will identify the potential mechanism by which viral factors dampen chemokine expression in infected IECs. Collectively, we expect these studies to establish a new paradigm of mucosal antiviral immunity, especially early in life when most enteric infections occur. We also expect to identify novel rotavirus immune evasion strategies, which will inform new strategies to develop host- based broad-spectrum antiviral therapeutics and next-generation vaccine candidates.

项目摘要 小肠肠上皮细胞（IEC）是针对人类肠道病毒的第一道防线， 婴儿和幼儿的腹泻和死亡的最常见和主要原因。 IECS 与小肠中的上皮内淋巴细胞（IEL）进行交流，并编排抗病毒反应 大量研究了。我们的总体目标是更好地定义宿主免疫信号通路 感染过程中的胃肠道，并使用该信息开发治疗干预措施 减轻腹泻和后遗症。 我们的初步结果与我们的期望形成鲜明对比，并证明了两个IEL子集的数量 在体内轮状病毒感染期间，大大降低了。我们还发现轮状病毒感染改变了 感染的人和小鼠IEC中几种促炎性趋化因子的表达。基于这些数据 和先前的出版物，我们假设IEL是主持人防御的重要调解人 轮状病毒感染和轮状病毒编码的因素拮抗IEL的抗病毒活性 通过抑制趋化因子表达来宿主小肠。 目前，缺乏合适的模型系统来妨碍测试这些假设。因此，我们 已经开发了一种高度可牵引的鼠轮状病毒反向遗传学方法，一种病理相关的新生儿 小鼠模型以及几个创新的原代人和鼠小肠癌培养系统， 这将为IEC-EIL串扰的理解提供前所未有的解决方案 病毒感染。 在AIM 1中，我们将使用免疫学在体内定义IELS在轮状病毒感染中的功能性抗病毒作用 接近和基因敲除小鼠。在AIM 2中，我们将确定病毒因素的潜在机制 受感染的IEC中的趋化因子表达。总体而言，我们希望这些研究能够建立一个新的 粘膜抗病毒免疫的范式，尤其是在大多数肠道感染发生的早期。我们也是 期望确定新型轮状病毒免疫逃避策略，这将为发展宿主的新策略提供信息 - 基于广谱的抗病毒治疗和下一代疫苗候选者。