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: 9980289
• 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/9980289
• 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 细胞间的相互作用对于免疫反应的特异性至关重要。然而，一种方法是 直接测量这些相互作用仍然缺乏，这为该领域创造了一个盲点。我的目标是开发一种 监测这种体外和体内相互作用的大范围系统，具有分子特异性(即，包括 关于涉及哪些受体/配体的信息)和在单个细胞水平上。这个系统，我们称之为 通过分类标记细胞间接触标记免疫伙伴关系(LIPSTIC)，使用酶标记 作为一种“细胞口红”跨越免疫突触，能够标记经历了 另一组细胞带有可通过流式细胞仪或显微镜检测到的标记。我们最初的证据是- 原理实验表明，该系统对CD40L-CD40在体外和体内的相互作用都有很好的效果。 活着。在先锋奖下，我建议将LIPSTIC技术扩展到多个受体配体，并 其他模式的标记和实验方法，因此它对免疫学家在 将军。这将在研究抗原启动T细胞反应的科学背景下进行 递呈细胞(APC)。我们将(I)扩展LIPSTIC调色板以标记多个受体之间的相互作用- 参与T细胞-APC串扰的配基对；(Ii)建立用于组合标记的双色系统 涉及多个受体-配体对或细胞群体的相互作用；以及(Iii)使用LIPSTIC作为工具 从全基因组文库中鉴定TCR配体。我们希望沿着这些方向发展LIPSTIC LINES有可能使其成为免疫学家的关键工具，同时提供对T细胞生物学的洞察 体内引爆，这是任何其他方法都无法获得的。