Modeling Shigella Interaction with Innate Cells in Enteroid Co-Cultures to Inform Vaccine Development

模拟肠类共培养物中志贺氏菌与先天细胞的相互作用，为疫苗开发提供信息

• 批准号: 10190303
• 负责人: Eileen M. Barry
• 金额: $ 37.71万
• 依托单位: UNIVERSITY OF MARYLAND BALTIMORE
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-07-01 至 2026-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10190303
• 关键词: 5 year old; Animal Model; Attenuated; Attenuated Vaccines; Biopsy; Cells; Cessation of life; Characteristics; Child; Clinical; Coculture Techniques; Complex; Dendritic Cells; Dendritic cell activation; Developing Countries; Diarrhea; Disease; Disease Progression; Dysentery; Endemic Diseases; Engraftment; Epithelial; Escherichia coli; Evaluation; Event; Funding; Gastrointestinal Physiology; Health Priorities; Human; Immune; Immune response; Infection; Inflammatory Response; Innate Immune Response; Intervention; Kinetics; Lymphocyte; M cell; Modeling; Multi-Drug Resistance; Mutation; Pathogenesis; Pathogenicity; Patients; Physiological; Population; Process; Series; Shigella; Shigella Infections; Shigella Vaccines; Shigella flexneri; Shigella sonnei; Symptoms; System; Therapeutic Intervention; Virulence Factors; burden of illness; gastrointestinal; global health; immune activation; insight; intestinal epithelium; intraepithelial; macrophage; novel; pathogen; preventive intervention; response; synergism; therapy development; tool; transcytosis; uptake; vaccine acceptance; vaccine candidate; vaccine development; vaccine evaluation

Project 2: Modeling Shigella Interaction with Innate Cells in Enteroid Co-Cultures to Inform Vaccine Development Shigella is responsible for a significant burden of disease in multiple populations within the US and worldwide causing an estimated 163 million cases and >74,000 deaths per year. The greatest impact is in children under 5 years of age in developing countries where Shigella was identified as the most important pathogen causing diarrhea in 12-59 month old children. The widespread isolation of multiple drug resistant isolates limiting therapeutic interventions and the continued high levels of endemic disease underscore the significance of Shigella as a global health priority and reinforce the need for preventative interventions and vaccine development. Infection with Shigella results in diarrhea and dysentery following transcytosis of the epithelial barrier, invasion of gastrointestinal cells, intracellular replication and induction of a severe inflammatory response. Interaction with host innate immune cells including macrophages (MΦ), dendritic cells (DCs) and intraepithelial lymphocytes (IEL) are critical events in progression of disease symptoms as well as in initiation of a protective immune response. The human enteroid model provides a highly human relevant multicellular system that recapitulates important aspects of gastrointestinal physiology. Advances during the first P01 funding cycle included the addition of M cells and immune cell co-culture in enteroids. This complex model provides a system in which critical features of Shigella uptake, transcytosis, and engagement with immune cells can be identified which may serve as targets for interventional strategies. Using a series of isogenic Shigella strains with mutations in key virulence factors, we aim to define bacterial requirements for each stage of host engagement. Furthermore, the evaluation of a series of live attenuated vaccine strains that have been studied clinically will serve as tools to further understand bacterial requirements for the pathogenic process and allow determination of the utility of this model to distinguish promising vaccine candidates. Building on our current capability of M cell incorporation, we will use the M cell enteroid model to characterize sequential stages of Shigella-host interactions including the process of uptake and transcytosis, and subsequent interaction with critical innate immune cells. These studies will reveal novel aspects of host-pathogen interactions and ensuing innate immune responses to this pathogen that better reflect what occurs in humans; new insights will be provided that will guide intervention strategies.

项目2：建模志贺氏菌与肠道共培养中的先天细胞相互作用以告知疫苗 发展 Shigella负责美国和全球多个人口中的疾病大量伯恩。 估计每年造成1.63亿例病例和> 74,000例死亡。最大的影响是儿童 志贺氏菌被确定为最重要的病原体的发展中国家5岁以下 在12-59个月大的孩子中引起腹泻。多种抗药性分离株的宽度分离 限制治疗干预措施和持续高水平的内在疾病强调了 志贺氏菌作为全球卫生优先事项的重要性，并加强了预防干预措施的需求和 疫苗开发。志贺氏菌的感染导致腹泻和痢疾后脑介绍 上皮屏障，胃肠细胞的侵袭，细胞内复制和诱导严重 炎症反应。与宿主先天免疫细胞的相互作用，包括巨噬细胞（Mφ），树突状 细胞（DC）和上皮内淋巴细胞（IEL）是疾病症状进展的关键事件 以及受保护的免疫反应的启动。人类肠模型提供了高度人类 相关的多细胞系统概括了胃肠道生理学的重要方面。进步 在第一个P01融资周期中，包括在肠动物中添加M细胞和免疫细胞共培养。 这个复杂的模型提供了一个系统，其中志贺氏菌吸收，跨胞菌和 可以识别与免疫细胞的互动，这可以作为介入策略的目标。 使用一系列与关键病毒因子突变的等生志菌菌株，我们旨在定义细菌 主机参与的每个阶段的要求。此外，评估一系列活跃的衰减 在临床上研究的疫苗菌株将用作进一步了解细菌的工具 病原过程的要求，并允许确定该模型的效用以区分 有希望的疫苗候选者。以我们当前的M细胞结合能力为基础，我们将使用M 细胞肠模型以表征志贺氏菌 - 宿主相互作用的顺序阶段，包括 摄取和跨胞菌病，以及随后与关键先天免疫细胞的相互作用。这些研究会 揭示宿主病原体相互作用的新颖方面并确保对这种病原体的先天免疫反应 这样可以更好地反映人类发生的事情；将提供新的见解，以指导干预策略。