Bacterial-mediated enhancement of Coxsackievirus

细菌介导的柯萨奇病毒增强

• 批准号: 10634743
• 负责人: Christopher Michael Robinson
• 金额: $ 48.58万
• 依托单位: INDIANA UNIV-PURDUE UNIV AT INDIANAPOLIS
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-01 至 2027-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10634743
• 关键词: Adult; Bacteria; Binding; Binding Sites; Cell Wall; Child; Communities; Coxsackie Viruses; Coxsackievirus Infections; Cryoelectron Microscopy; Data; Development; Elements; Escherichia coli; Foot Diseases; Future; Gastrointestinal tract structure; Goals; Hand' s disease; Home; In Vitro; Infection; Lipopolysaccharides; Mediating; Meningoencephalitis; Mouth Diseases; Oral; Pathogenesis; Prevention; Route; Salmonella; Salmonella enterica; Structure; Viral; Viral Pathogenesis; Virion; Virus Replication; human pathogen; in vivo; microbiota; mouse model; mutant; novel therapeutics; thermostability; transmission process; viral myocarditis

Project Summary Coxsackievirus is a common infection that causes viral myocarditis, meningoencephalitis, and hand, foot, and mouth disease in children and adults. Coxsackievirus is transmitted by the fecal-oral route and initiates infection in the gastrointestinal tract. Currently, there is a lack of effective prevention and treatment options for Coxsackievirus infections. Using a mouse model, recent data demonstrate that Coxsackievirus B3 (CVB3) utilizes bacteria to aid viral replication and pathogenesis in vivo. However, the mechanism for this observation is unclear. In preliminary data, we have demonstrated that bacteria can enhance CVB3 infectivity and thermostability in vitro. We also found that Gram-negative Salmonella enterica binds to CVB3 and can enhance infectivity and stability of CVB3 while Escherichia coli does not. We also found that structures in the major cell wall component of Salmonella, lipopolysaccharide (LPS), can impact viral stability. These data suggest that specific bacteria and bacterial cell wall components are required to enhance CVB3. Therefore, this study aims to elucidate the bacterial structure and site of binding to CVB3, and investigate the mechanisms by which bacteria aid viral thermostability. This goal will be accomplished in three aims. In Aim 1, we will determine the bacterial cell wall components that interact with CVB3. Using Salmonella LPS mutants, we will characterize the critical elements of LPS required to interact with CVB3. In Aim 2, we will identify the virion binding site between bacteria and Coxsackievirus. Using a combination of state-of-the-art cryogenic electron microscopy and CVB3 mutants, we will elucidate the LPS footprint on the CVB3 virion. Finally, in Aim 3, we will determine the effect of lipopolysaccharide on CVB3 infectivity and viral stability. Overall, this project will significantly impact our understanding of viral-microflora interactions and aid future studies to identify antiviral targets for the development of novel therapeutics.

项目摘要 柯萨奇病毒是一种常见的感染，可引起病毒性心肌炎，脑膜脑炎，手足部和 儿童和成人的口腔疾病。柯萨奇病毒通过粪便-口腔途径传播，并引发感染 在胃肠道里。目前，缺乏有效的预防和治疗方案 柯萨奇病毒感染。利用小鼠模型，最近的数据表明柯萨奇病毒B3(CVB3) 利用细菌在体内帮助病毒复制和致病。然而，这种观察的机制是 不清楚。在初步数据中，我们已经证明细菌可以增强CVB3的传染性和 体外热稳定性。我们还发现革兰氏阴性沙门氏菌与CVB3结合，并能增强 CVB3的感染性和稳定性，而大肠杆菌不具有。我们还发现，主细胞中的结构 沙门氏菌的壁组分--脂多糖(LPS)会影响病毒的稳定性。这些数据表明 需要特定的细菌和细菌细胞壁成分来增强CVB3。因此，本研究旨在 阐明与CVB3结合的细菌结构和部位，并探讨细菌与CVB3结合的机制。 有助于病毒的热稳定性。这一目标将分三个目标实现。在目标1中，我们将确定细菌 与CVB3相互作用的细胞壁成分。使用沙门氏菌内毒素突变体，我们将描述关键的 与CVB3相互作用所需的内毒素元件。在目标2中，我们将确定细菌之间的病毒粒子结合部位。 和柯萨奇病毒。使用最先进的低温电子显微镜和CVB3突变体的组合， 我们将阐明内毒素在CVB3病毒粒子上的足迹。最后，在目标3中，我们将确定 脂多糖对CVB3感染性和病毒稳定性的影响。总体而言，这个项目将对我们的 了解病毒-微生物区系的相互作用，并帮助未来的研究确定抗病毒靶点 开发新的治疗方法。