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: 10686834
• 负责人: Eileen M. Barry
• 金额: $ 38.45万
• 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/10686834
• 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：模拟志贺氏菌与肠共培养物中先天细胞的相互作用，以告知疫苗 发展 志贺氏菌是美国和世界范围内多个人群的重大疾病负担 估计每年造成1.63亿例病例和> 74，000例死亡。影响最大的是儿童 在志贺氏菌被确定为最重要病原体的发展中国家， 导致12-59个月大的儿童腹泻。多重耐药菌株的广泛分离 有限的治疗干预措施和持续的高水平的地方病强调， 志贺氏菌作为全球卫生优先事项的重要性，并加强预防干预的必要性， 疫苗研发。志贺氏菌感染导致腹泻和痢疾后转胞吞的 上皮屏障，胃肠道细胞的侵袭，细胞内复制和诱导严重的 炎症反应。与宿主固有免疫细胞（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）是疾病症状进展中的关键事件， 以及启动保护性免疫反应。人类肠模型提供了高度人源化的 相关的多细胞系统，概括了胃肠道生理学的重要方面。进展 在第一个P01资助周期期间，包括在肠样组织中添加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，其中志贺氏菌的摄取，转胞吞作用， 可以鉴定与免疫细胞的接合，其可以作为干预策略的靶。 利用一系列关键毒力因子突变的志贺菌等基因菌株，我们的目的是确定细菌 主机参与的每个阶段的要求。此外，还对一系列减毒活疫苗进行了评价， 已经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实用性，以区分 有希望的候选疫苗基于我们目前的M细胞掺入能力，我们将使用M 细胞类肠模型来表征志贺氏菌-宿主相互作用的连续阶段，包括 摄取和转胞吞作用，以及随后与关键的先天免疫细胞的相互作用。这些研究将 揭示了宿主-病原体相互作用的新方面，以及随之而来的对该病原体的先天免疫应答 更好地反映人类发生的情况;将提供新的见解，指导干预战略。