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.

项目总结-核心c