Immunoassay and Imaging Core

免疫测定和成像核心

• 批准号: 10594526
• 负责人: Stuart P Weisberg
• 金额: $ 22.66万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-01-01 至 2027-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10594526
• 关键词: 2019-nCoV; Affinity; Age; Antibodies; Antibody Response; Antibody Specificity; Antibody titer measurement; Antigen Presentation; Antigen-Antibody Complex; Antigens; Area; Autoimmune Diseases; B-Cell Antigen Receptor; B-Lymphocytes; Biological Assay; Blood; Cell Communication; Cells; Cellular Assay; Cellular Immunity; Circulation; Complement; Data; Data Set; Dendritic Cells; Detection; Development; Dimensions; Exposure to; Gene Expression Profile; Generations; Goals; Helper-Inducer T-Lymphocyte; Human; Humoral Immunities; Image; Immune; Immune response; Immunity; Immunoassay; Immunologic Memory; Immunology procedure; Immunosuppression; In Situ; Infection; Influenza; Influenza Hemagglutinin; Invaded; Location; Lung; Lymphocyte; Lymphoid; Lymphoid Tissue; Macrophage; Maintenance; Mediating; Molecular Profiling; Nucleocapsid; Organ Donor; Pharmaceutical Preparations; Population; Process; Reaction; Regulatory T-Lymphocyte; SARS-CoV-2 antibody; Serology; Serology test; Serum; Services; Shapes; Site; Stains; Structure of germinal center of lymph node; Surface; System; T cell response; T memory cell; T-Lymphocyte; Therapeutic immunosuppression; Tissue Preservation; Tissues; Vaccination; Vaccine Antigen; Validation; Viral; Viral Antibodies; Virus; Virus Diseases; Visualization; adaptive immune response; anti-influenza; antigen-specific T cells; arm; biomarker panel; cell type; cohort; data modeling; density; design; effector T cell; experience; human tissue; imaging platform; immune imaging; influenza virus vaccine; liquid crystal polymer; lymphatic vessel; lymphocyte trafficking; multiple sclerosis patient; multiplexed imaging; neutralizing antibody; novel; novel marker; organizational structure; pathogen; pathogenic virus; post SARS-CoV-2 infection; programs; receptor binding; response; single cell sequencing; tissue resident memory T cell; vaccine response

IMMUNOASSAY AND IMAGING CORE: PROJECT SUMMARY Key correlates of immune protection against viral pathogens include virus specific memory T cells resident in tissues and high affinity virus-neutralizing antibodies circulating in blood. During a primary immune response, newly encountered antigen is transported from the sites of exposure to regional lymphoid tissue where naive lymphocytes encounter antigen-presenting dendritic cells and macrophages which critically regulate the priming, expansion and activation of antigen specific T and B cells. Memory T cells derive from activated or effector T cells generated during the primary immune response and differentiate into circulating, tissue surveilling memory T cells as well as tissue resident memory T cells (TRM) that are retained in diverse tissue sites. Immune protection at barrier tissue sites – the primary point of pathogen invasion such as lung - is mediated by TRM established by previous antigen exposures that are specially adapted to persist in the tissue microenvironment and provide a highly localized defense against pathogen re-entry. Long lasting humoral immunity is established via selection of somatically hypermutated B cell antigen receptors in the germinal centers of lymphoid tissue – a process dependent on help from CD4 T follicular helper cells. Thus development and maintenance of cellular and humoral immunity is critically dependent on immune cell interactions in lymphoid and non-lymphoid barrier tissues. This service core will provide quantitative assays of the cellular and humoral arms of the immune response using in situ multiplex staining to define key T cell and dendritic cell populations mediating immune protection in tissues and serological assays to define humoral anti-SARS-CoV- 2 and anti-influenza antibody responses to infection and vaccination. The Aims of the service core are 1) Perform large scale quantitative spatial analysis of immune cells within human tissues; and 2) Provide quantitative and functional analysis of anti-viral antibodies in human serum. For Aim 1, molecular signatures obtained by single cell sequencing of antigen-specific T cells (Project 1) and dendritic cells (Project 2) in human tissues will be used to build multiplex imaging panels to visualize these cells in-situ. The core will support tissue preservation, multiplex staining and analysis of multiplex imaging with a focus on identifying novel antigen-specific TRM and dendritic cell populations across multiple lymphoid and barrier tissue sites and donors. These studies will complement detailed molecular signatures of tissue immune cells with information about their spatial context and microenvironment. For Aim 2, assays of circulating virus specific antibodies will be used to assess SARS-CoV-2 immune protection and vaccination status in organ donors for correlation with tissue immune responses and other donor-specific covariates (Project 1) and to assess SARS-CoV-2 and influenza vaccine responses longitudinally in multiple sclerosis patients taking immunosuppressive medications (Project 3). Together, data generated from this core, in support of the overall program goals, will help define factors that shape tissue and humoral immune protection in humans.

免疫测定和成像核心：项目摘要 免疫学保护病毒病原体的关键相关性包括病毒特异性记忆T细胞居民 组织和高亲和力病毒中和化抗体在血液中循环。在初级免疫反应期间， 新遇到的抗原从暴露于区域淋巴组织的部位中运输 淋巴细胞会遇到抗原呈递的树突状细胞和巨噬细胞，这些细胞和巨噬细胞严格调节 抗原特异性T和B细胞的启动，膨胀和激活。内存T细胞来自激活或 在初级免疫反应过程中产生的效应T细胞并分化为循环，组织 监视记忆T细胞以及保留在潜水组织中的组织居民记忆T细胞（TRM） 站点。屏障组织部位的免疫保护 - 病原体入侵（例如肺）的主要点 - 由以前的抗原暴露建立的TRM介导的，这些暴露是专门适应在组织中的 微环境，并提供高度局部的防御病原体重新进入。持久的幽默 通过选择生发中的体形高水沉积的B细胞抗原受体建立免疫力 淋巴组织中心 - 取决于CD4 T卵泡辅助细胞帮助的过程。那个发展 细胞和体液免疫史的维持至关重要取决于免疫细胞相互作用 淋巴样和非淋巴屏障组织。该服务核心将提供细胞和细胞的定量评估 免疫反应的体液臂使用原位多重染色来定义关键T细胞和树突状细胞 介导组织中免疫保护和血清学测定的种群，以定义体液抗SARS-COV- 2和对感染和疫苗接种的抗Imfluenza抗体反应。服务核心的目的是1） 对人体组织中免疫细胞进行大规模定量空间分析； 2）提供 人血清中抗病毒抗体的定量和功能分析。对于AIM 1，分子特征 通过抗原特异性T细胞（项目1）和树突状细胞（项目2）的单细胞测序获得 人体组织将用于构建多重成像面板，以可视化这些细胞的原位。核心意愿 支持组织制备，多次染色和对多个成像的分析，重点是识别 跨多个淋巴机和屏障组织部位的新型抗原特异性TRM和树突状细胞群 捐助者。这些研究将补充组织免疫细胞的详细分子特征 关于它们的空间环境和微环境。对于AIM 2，循环病毒特异性抗体的测定将 用于评估器官捐赠者中的SARS-COV-2免疫保护和疫苗接种状态，以与 组织免疫复杂和其他供体特异性协变量（项目1），并评估SARS-COV-2和 对服用免疫抑制药物的多发性硬化症患者的影响力疫苗反应纵向 （项目3）。从该核心生成的数据共同支持总体计划目标，将有助于定义 塑造人类组织和人类免疫保护的因素。