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: 8855411
• 负责人: Sean Curtis Bendall
• 金额: $ 26.58万
• 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/8855411
• 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 访问GI（高级共文化工程和肠道免疫学的单细胞统计）核心 C将为斯坦福大学的调查员提供一系列最前沿的方法（项目1、2和 飞行员）分析肠/胃上皮细胞及其与免疫细胞与与免疫细胞的相互作用 没有感染。与类器官B相关，我们将进行开发工作 产生整合粘膜免疫细胞的器官。具体来说，在AIM 1中，我们将高度利用 多路复用的单细胞分析（质量细胞仪）和成像（多路复用离子束成像 - MIBI） 肖恩·本德尔（Sean Bendall）在斯坦福大学开发的方式为项目提供细胞分析分析 跨越名称中心（项目1、2）。这些评估将是独特的，允许 人肠癌结构的简单表征，免疫细胞的整合和影响 病原体对细胞行为的影响。除此之外，核心C将有助于访问其他分析方式 可通过斯坦福大学的人类免疫监测核心（HIMC）获得，其中包括63 plex 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 流体化生物标志物用于单细胞QPCR分析。核心将利用他的团队Sean Bendall的专业知识 以及用于分析这些平台生成的数据以及适当的多平台数据的数据 在AIM 2中，Sarah Heilshorn和她的团队将为纳入调查人员提供新颖的测定 （项目1，2），使用允许研究细胞/细胞和细胞/病原体相互作用的微流体设备 高度控制的微环境。在AIM 3中，伊丽莎白·梅林斯（Elizabeth Mellins）和她的团队与 器官核心将为项目1提供类似M细胞的类器官，并表达II类并发展 具有M细胞生理水平的器官的产生。她的团队还将提供同质的免疫细胞 在被感染的情况下，向名称的研究者进行了针对免疫反应性的实验 肠癌（项目1）以及选择抗原呈现细胞与 未感染和感染的类器官。后者将与 器官核，开发人肠癌，更紧密地复制生理 肠道感染期间病原体遇到的微环境。