Determinants of Enteric Calicivirus Infection.

肠道杯状病毒感染的决定因素。

• 批准号: 10213588
• 负责人: TIBOR FARKAS
• 金额: $ 37.32万
• 依托单位: TEXAS A&M AGRILIFE RESEARCH
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-07-01 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10213588
• 关键词: Acute; Address; Apical; B-Lymphocytes; Bile fluid; Binding Sites; Biodiversity; Biological; Biological Models; Biopsy; Blood Group Antigens; CRISPR/Cas technology; Calicivirus; Calicivirus Infections; Cell Culture System; Cell Culture Techniques; Cell Line; Cell surface; Cessation of life; Consumption; Coxsackie Viruses; Data; Development; Diarrhea; Disease; Enteral; Enterobacter; Enterocytes; Environment; Epithelial; Exhibits; Extracellular Domain; Fever; Gastroenteritis; Genetic; Goals; Healthcare; Hospitalization; Human; In Vitro; Infection; Integration Host Factors; Intervention; Intestines; Laboratories; Lamina Propria; Lead; M cell; Macaca; Maps; Mediating; Modeling; Norovirus; Pathway interactions; Peripheral Blood Mononuclear Cell; Play; Positioning Attribute; Predisposition; Prevention strategy; Primates; Protein Isoforms; Public Health; Reproducibility; Research; Rhesus; Role; Serotyping; Site; Surface; System; Testing; Tight Junctions; Time; United States; Viral; Virus; Work; Zoonoses; adenovirus receptor; antigen binding; base; bile salts; comparative; cross-species transmission; enteric virus infection; genome wide screen; glycosylation; improved; infected B cell; insight; intestinal epithelium; monolayer; mutant; nonhuman primate; novel; permissiveness; polarized cell; preventive intervention; receptor; societal costs

PROJECT SUMMARY Human norovirus (HuNoV) gastroenteritis is a significant public health burden worldwide. The lack of a robust and reproducible HuNoV cell culture system still limits our ability to study fundamental aspects of HuNoV infection. While several recent breakthroughs increased our ability to propagate HuNoVs in vitro (e.g. B cell and human enteroid cultures), these systems are not robust enough, can be time consuming and expensive or hard to replicate. The surrogate models available for HuNoV research do not necessarily reflect essential biological features of HuNoVs and their natural host. In this proposal we will use our novel rhesus enteric caliciviruses (ReCV) model that reflects the biological features and diversity of HuNoVs to identify host determinants of infection. Zoonotic/interspecies transmission of ReCVs and HuNoVs between human and non- human primate hosts suggests evolutionary conservation of shared factors of host susceptibility between the two genera. Here we seek to identify host determinants of enteric calicivirus infection using CRISPR-Cas9 genome wide screening. We recently identified a functional ReCV entry receptor that is necessary for ReCV permissiveness in cell culture. We hypothesize that the ReCV entry receptor is also involved in HuNoV infections. This hypothesis will be tested in the following specific aims. Aim 1. Characterize receptor mediated ReCV infection. Aim 2: Comparative characterization of HuNoV and ReCV infections in enteroid and B cell cultures. In the first aim we will dissect the role of HBGAs in ReCV receptor mediated entry by using ReCV isolates with disctinct HBGA binding, cell lines expressing different HBGAs and/or the receptor, Enterobacter SENG-6 EPS and different synthetic HBGAs. We will also evaluate the role of the different transmembrane isoforms of the receptor in infection and map the ReCV interaction site and importance of glycosylation. In the second aim, we will evaluate the role of ReCV entry receptor in HuNoV infections in enteroid and B cell cultures and identify other cell surface components playing a role in HuNoV infections. We will also evaluate the mechanism of bile or bile salts in promoting HuNoV infections and the role of M cells in infections of polarized cells. Our long-term goal is to identify and characterize viral and host determinants of ReCV and HuNoV infections and to develop novel intervention/prevention strategies. Our proposal uses a novel enteric calicivirus model to understand viral entry and the role of bile and HBGAs in enteric viral infections. Our findings with ReCVs may be directly transferable to HuNoV infection. The major significance of this project is the identification of determinants of susceptibility to enteric calicivirus infection that can lead to improved HuNoV cell culture systems and new intervention/prevention strategies.

项目摘要 人类诺如病毒（Hunov）肠胃炎是全球的​​重大公共卫生负担。缺乏强大的 可再现的Hunov细胞培养系统仍然限制了我们研究Hunov基本方面的能力 感染。虽然最近的几个突破增加了我们在体外传播Hunovs的能力（例如B细胞 和人类肠道培养物），这些系统不够强大，可能会耗时且昂贵或 很难复制。可用于Hunov研究的替代模型不一定反映出必不可少的 霍诺夫及其天然宿主的生物特征。在这个建议中，我们将使用我们的小说恒河馆肠 蜡膜病毒（RECV）模型，反映了匈奴的生物学特征和多样性以识别宿主 感染的决定因素。人畜共动/种间传播RECV和人类和非人之间的Hunovs 人类灵长类动物主持人提出了对宿主易感性共享因素的进化保护 两个属。在这里，我们试图使用CRISPR-CAS9识别肠胃病毒感染的宿主决定因素 基因组广泛的筛查。我们最近确定了RECV所需的功能性RECV进入受体 细胞培养的允许性。我们假设RECV进入受体也参与了Hunov 感染。该假设将在以下特定目的中进行检验。 AIM 1。表征受体介导的 RECV感染。 AIM 2：Enteroid和B细胞中Hunov和RECV感染的比较表征 文化。在第一个目的中，我们将使用RECV剖析HBGA在RECV受体介导的进入中的作用 脱离HBGA结合的分离株，表达不同HBGA和/或受体的细胞系，肠杆菌 Seng-6 EP和不同的合成HBGA。我们还将评估不同跨膜的作用 受体在感染中的同工型并绘制RECV相互作用位点和糖基化的重要性。在 第二个目的，我们将评估RECV进入受体在霍诺夫感染中的作用 培养并鉴定其他细胞表面成分在Hunov感染中起作用。我们还将评估 胆汁或胆汁盐在促进Hunov感染中的机制以及M细胞在感染中的作用 偏振细胞。我们的长期目标是识别和表征RECV的病毒和宿主决定因素和 匈奴感染并制定新颖的干预/预防策略。我们的建议使用一个新颖的企业 彩色病毒模型了解病毒进入以及胆汁和HBGA在肠道病毒感染中的作用。我们的 带有RECV的发现可以直接转移到匈奴感染。该项目的主要意义是 鉴定对肠性膜病毒感染的易感性的决定因素，可以改善 Hunov细胞培养系统和新的干预/预防策略。