Core C: Advanced Co-Culture Engineering and Single Cell Statistics of Gut Immunology

核心C：肠道免疫学的高级共培养工程和单细胞统计

• 批准号: 9022402
• 负责人: Sean Curtis Bendall
• 金额: $ 24.97万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/9022402
• 关键词: Address; Antibodies; Antigen-Presenting Cells; Antigens; B-Lymphocytes; Behavior; Biochemical; Biocompatible Materials; Bioinformatics; Biological Assay; Biological Models; Blood; Cell Count; Cell Cycle; Cell Differentiation process; Cell Line; Cells; Cellular Structures; Chemotaxis; Coculture Techniques; Collaborations; Culture Techniques; Cytometry; Data; Data Analyses; Dendritic Cells; Development; Devices; Disease; Distal; Engineering; Enteral; Environment; Enzyme-Linked Immunosorbent Assay; Epithelial Cells; Epithelium; Foundations; Generations; Goals; Helicobacter pylori; Human; Image; Image Cytometry; Immune; Immunologic Monitoring; Immunology; Individual; Infection; Interferons; Intestines; Investigation; Ions; Isotopes; Ligands; M cell; MHC Class II Genes; Measurement; Methods; Microfluidic Microchips; Microfluidics; Modality; Monitor; Organoids; Pathway interactions; Phenotype; Physiological; Population; Predisposition; Production; Raji Cell; Reagent; Research; Research Personnel; Rotavirus; Salmonella; Salmonella typhi; Stomach; Structure; System; TNFSF11 gene; Technology; Time; Tissue Engineering; Tissues; Work; base; cell type; chemokine; cytokine; data integration; design; gastrointestinal epithelium; in vivo; intercellular communication; macrophage; migration; monocyte; novel; optical imaging; pathogen; research study; response; single cell analysis; statistics; trafficking

PROJECT SUMMARY – CORE C The ACCESS-GI (Advanced Co-culture Engineering and Single cell Statistics of Gut Immunology) Core C will provide a range of cutting edge approaches to Stanford NAMSED investigators (Projects 1, 2 and pilots) to analyze intestinal/gastric epithelial cells and their interactions with immune cells both with and without infection. In association with the Organoid Core B, we will carry out development work towards the generation of organoids that integrate mucosal immune cells. Specifically, in Aim 1, we will leverage highly multiplexed, single cells analysis (CyTOF mass cytometry) and imaging (multiplexed ion beam imaging – MIBI) modalities collaboratively developed at Stanford by Sean Bendall to provide cell analysis assays for projects across the NAMSED center (Projects 1, 2). These assays will be uniquely beneficial, allowing the simultaneous characterization of human gut organoid structure, the integration of immune cells, and the effects of pathogens on cellular behavior. Further to this, Core C will facilitate access to other analysis modalities available through Stanford's human immune monitoring core (HIMC), including 63-plex Luminex for quantitative measurement of human cytokines and chemokines, standard ELISA for non-Luminex targets, and Fluidigm Biomark for single cell qPCR analysis. The core will leverage the expertise of Sean Bendall, his team, and the HIMC for analysis of data generated by these platforms and, where appropriate, for multi-platform data integration. In Aim 2, Sarah Heilshorn and her team will provide novel assays to NAMSED investigators (Projects 1, 2), using microfluidic devices that allow investigation of cell/cell and cell/pathogen interactions in highly controlled micro-environments. In Aim 3, Elizabeth Mellins and her team, in collaboration with the Organoid Core, will provide M-cell-like organoids to Project 1, and class II expressing organoids and develop production of organoids with physiologic levels of M cells. Her team will also provide homogenous immune cell populations to NAMSED investigators for experiments addressing immune reactivity in the context of infected intestinal organoids (Project 1) as well as study interactions of selected antigen presenting cells with uninfected and infected organoids. The latter will form the foundation of a key effort, in collaboration with the Organoid Core, to develop human intestinal organoids that more closely replicate the physiologic microenvironment encountered by pathogens during enteric infection.

项目摘要 – 核心 C ACCESS-GI（肠道免疫学的高级共培养工程和单细胞统计）核心 C 将为斯坦福大学 NAMSED 研究人员提供一系列前沿方法（项目 1、2 和 飞行员）分析肠/胃上皮细胞及其与免疫细胞的相互作用 我们将与Organoid Core B一起进行开发工作，以实现无感染。 具体来说，在目标 1 中，我们将利用整合粘膜高度免疫细胞的类器官。 多重、单细胞分析（CyTOF 质谱流式分析）和成像（多重离子束成像 – MIBI） 由 Sean Bendall 在斯坦福大学合作开发的模式，为项目提供细胞分析检测 整个 NAMSED 中心（项目 1、2）将具有独特的优势，使 同时表征人​​类肠道类器官结构、免疫细胞的整合及其影响 除此之外，Core C 将促进其他分析方式的使用。 可通过斯坦福大学的人体免疫监测核心 (HIMC) 获得，包括 63 重 Luminex 人类细胞因子和趋化因子的定量测量，非 Luminex 靶标的标准 ELISA，以及 用于单细胞 qPCR 分析的 Fluidigm Biomark 核心将利用 Sean Bendall 团队的专业知识， HIMC 用于分析这些平台生成的数据，并在适当情况下分析多平台数据 在目标 2 中，Sarah Heilshorn 和她的团队将为 NAMSED 研究人员提供新颖的检测方法。 （项目 1、2），使用微流体装置来研究细胞/细胞和细胞/病原体之间的相互作用 在《目标 3》中，伊丽莎白·梅林斯 (Elizabeth Mellins) 和她的团队与 Organoid Core，将为项目1提供M细胞样类器官，以及II类表达类器官并开发 她的团队还将生产具有生理水平的 M 细胞的类器官。 向 NAMSED 研究人员提供有关感染者免疫反应的实验 肠道类器官（项目 1）以及研究选定的抗原呈递细胞与 未感染和感染的类器官将构成与该组织合作的关键工作的基础。 类器官核心，开发更接近地复制生理学的人类肠道类器官 肠道感染过程中病原体遇到的微环境。