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: 10745566
• 负责人: Eileen M. Barry
• 金额: $ 2.59万
• 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/10745566
• 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; Epithelium; Escherichia coli; Evaluation; Event; Funding; Gastrointestinal Physiology; Health Priorities; Human; Immune; Immune response; Infection; Inflammatory Response; Innate Immune Response; Intervention; Invaded; Kinetics; Lymphocyte; M cell; Macrophage; 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; 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：模拟肠类共培养物中志贺氏菌与先天细胞的相互作用，为疫苗提供信息 发展 志贺氏菌对美国和全世界多个人群造成重大疾病负担 估计每年造成 1.63 亿病例和超过 74,000 人死亡，其中影响最大的是儿童。 志贺氏菌被确定为最重要病原体的发展中国家 5 岁以下儿童 腹泻 引起 12-59 个月大儿童腹泻 广泛分离出多种耐药菌株。 治疗干预措施的限制和地方病的持续高水平凸显了 志贺氏菌作为全球卫生优先事项的重要性，并强化了预防性干预措施和 疫苗开发后，志贺氏菌感染会导致腹泻和痢疾。 上皮屏障、胃肠道细胞的侵袭、细胞内复制和诱导严重的 与宿主先天免疫细胞（包括巨噬细胞（MΦ）、树突状细胞）的相互作用。 细胞（DC）和上皮内淋巴细胞（IEL）是疾病症状进展的关键事件，如 以及启动保护性免疫反应，人类肠样模型提供了高度人性化的模型。 相关的多细胞系统概括了胃肠道生理学进展的重要方面。 在第一个 P01 资助周期中，包括在肠类中添加 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，其中志贺氏菌摄取、转胞吞作用和 可以确定与免疫细胞的相互作用，这可以作为干预策略的目标。 使用一系列关键毒力因子发生突变的同基因志贺氏菌菌株，我们的目标是定义细菌 此外，还有一系列现场衰减的评估。 已进行临床研究的疫苗株将作为进一步了解细菌的工具 致病过程的要求，并允许确定该模型的效用以区分 基于我们目前有前景的 M 细胞整合能力，我们将使用 M 细胞肠样模型，用于表征志贺氏菌与宿主相互作用的连续阶段，包括 摄取和转胞吞作用，以及随后与关键先天免疫细胞的相互作用。 揭示宿主-病原体相互作用的新方面以及随后对该病原体的先天免疫反应 更好地反映人类发生的情况；将提供指导干预策略的新见解。