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

项目2：在肠道共培养中模拟志贺氏菌与天然细胞的相互作用，为疫苗提供信息 发展 志贺氏菌在美国和世界各地的多个人群中造成了严重的疾病负担 据估计，每年造成1.63亿起病例和7.4万人死亡。对儿童的影响最大 发展中国家5岁以下儿童中志贺氏菌被确定为最重要的病原体 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地方病水平突显了 志贺氏菌作为全球卫生优先事项的重要性，并加强预防性干预和 疫苗研发。感染志贺氏菌导致腹泻和痢疾，继而发生细胞输血 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。与宿主天然免疫细胞的相互作用，包括巨噬细胞(MΦ)、树突状细胞 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。人类肠状骨模型提供了高度人性化的 相关的多细胞系统，概括胃肠生理学的重要方面。预付款 在第一个P01资助周期中，包括在肠样体中加入M细胞和免疫细胞共培养。 这一复杂的模型提供了一种系统，在该系统中志贺氏菌的摄取、跨细胞和 可以确定与免疫细胞的接触，这些细胞可能是干预策略的靶点。 利用一系列具有关键毒力因子突变的同源志贺氏菌，我们的目标是定义细菌 主持人参与的每个阶段的要求。此外，对一系列带电衰减的评价 经过临床研究的疫苗毒株将成为进一步了解细菌的工具。 对致病过程的要求，并允许测定此模型的实用性以区分 前景看好的候选疫苗。在我们目前M细胞掺入能力的基础上，我们将使用M 描述志贺氏菌-宿主相互作用的连续阶段的细胞肠样模型，包括 摄取和跨细胞作用，以及随后与关键的先天免疫细胞的相互作用。这些研究将 揭示宿主-病原体相互作用和随后对该病原体的先天免疫反应的新方面 这更好地反映了在人类身上发生的事情；将提供新的见解来指导干预策略。

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.