Quantifying Cell-Cell Interactions in the Immune System by Trans-Synaptic Labeling

通过跨突触标记量化免疫系统中的细胞间相互作用

• 批准号: 10213593
• 负责人: Gabriel D Victora
• 金额: $ 118.65万
• 依托单位: ROCKEFELLER UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-30 至 2023-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10213593
• 关键词: Antigen-Presenting Cells; Award; Biology; Cell Communication; Cells; Color; Communicable Diseases; Development; Flow Cytometry; Genomic Library; Goals; Immune; Immune system; Immunologist; Immunotherapy; In Vitro; Individual; Label; Ligands; Malignant Neoplasms; Measures; Methodology; Microscopy; Molecular; Monitor; Population; Public Health; Regimen; Research; Retinal blind spot; Specificity; Synapses; System; T cell response; T-Lymphocyte; TNFRSF5 gene; TNFSF5 gene; Technology; Work; combinatorial; experimental study; immunological synapse; immunoreaction; improved; in vivo; insight; novel; receptor; tool; vaccine development; whole genome

Project Summary Cell-cell interactions are essential for the specificity of immunological reactions. However, an approach to measuring these interactions directly is still lacking, creating a blind spot for the field. My goal is to develop a broad-ranging system to monitor such interactions in vitro and in vivo, with molecular specificity (i.e., including information on which receptors/ligands are involved) and at the individual cell level. This system, which we call Labeling Immune Partnerships by SorTagging Intercellular Contacts (LIPSTIC), uses enzymatic labeling across immune synapses as a “cellular lipstick” capable of marking cells that have undergone interactions with another cell population with labels that are detectable by flow cytometry or microscopy. Our initial proof-of- principle experiments show that this system works robustly for the CD40L-CD40 interaction, both in vitro and in vivo. Under the Pioneer award, I propose to expand the LIPSTIC technology to multiple receptor ligands and to other modes of labeling and experimental approaches, so that it becomes widely useful to immunologists in general. This will be done within the scientific context of studying the priming of T cell responses by antigen presenting cells (APCs). We will (i) extend the LIPSTIC palette to label interactions between multiple receptor- ligand pairs involved in T cell-APC cross-talk; (ii) develop dual-color systems for combinatorial labeling of interactions involving multiple receptor-ligand pairs or cell populations; and (iii) use LIPSTIC as a tool to identify TCR ligands from among whole-genome libraries. We expect that developing LIPSTIC along these lines has the potential to make it a key tool for immunologists, while providing insight into the biology of T cell priming in vivo that could not be obtained by any other means.

项目摘要 细胞 - 细胞相互作用对于免疫反应的特异性至关重要。但是，一种方法 仍然缺乏直接测量这些相互作用，为该领域创造了一个盲点。我的目标是开发 具有分子特异性的体外和体内相互作用的宽范围系统（即 有关涉及哪些受体/配体的信息）以及单个细胞水平。这个系统，我们称之为 通过分类细胞间触点（肥大）标记免疫合作伙伴关系，使用酶标记 跨免疫类吸收是一种“细胞唇膏”，能够标记与与之相互作用的细胞 另一个具有流式细胞仪或显微镜可检测的标签的细胞群。我们最初的证明 原理实验表明，该系统在体外和IN都适用于CD40L-CD40相互作用 体内。根据先驱奖，我建议将口头技术扩展到多个接收器配体，并将其扩展到 标签和实验方法的其他模式，以使其对免疫学家的广泛有用 一般的。这将在研究抗原启动T细胞反应的科学背景下完成 呈现细胞（APC）。我们将（i）将肥大的调色板扩展到多个接收器之间的相互作用 - 参与T细胞APC串扰的配体对； （ii）开发用于组合标记的双色系统 涉及多个接收器配体对或细胞群体的相互作用； （iii）使用肥大作为工具 从全基因组库中识别TCR配体。我们希望沿这些发展 线条有可能使其成为免疫学家的关键工具，同时洞悉T细胞的生物学 无法通过任何其他方式获得的体内启动。