Tissue regulation of T cell function - Reagents Core

T 细胞功能的组织调节 - 试剂核心

• 批准号: 10477319
• 负责人: JIM F Miller
• 金额: $ 30.78万
• 依托单位: UNIVERSITY OF ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-06-01 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10477319
• 关键词: Abbreviations; Animal Model; Animals; Antigen-Presenting Cells; Breeding; CD8B1 gene; Calcium; Cell physiology; Cells; Color; Cues; Cytoskeletal Proteins; Event; Extracellular Matrix; Fluorescence Resonance Energy Transfer; Funding; Gene Expression; Generations; Genetic; Goals; Image; Immune response; Immunologic Memory; Individual; Infection; Inflammatory Response; Integrins; Intracellular Membranes; Label; Leukocytes; LoxP-flanked allele; Maintenance; Memory; Molecular; Monitor; Mouse Strains; Mus; Myeloid Cells; Peripheral; Population; Positioning Attribute; Process; Proteins; Reagent; Reporter; Research Personnel; Research Project Grants; Retroviridae; Signal Transduction; Site; Structural Protein; T cell regulation; T-Cell Activation; T-Cell Receptor; T-Lymphocyte; Tissues; Transgenic Organisms; cell motility; cell type; effector T cell; experimental study; imaging modality; in vivo imaging; migration; mouse model; multiphoton microscopy; pathogen; programs; recruit; repository; response

PROJECT SUMMARY / ABSTRACT – REAGENT CORE Recruitment and localization of leukocytes to sites of infection in inflamed tissue is a dynamic and regulated process that depends on cross talk between different cell populations, responses to specific environmental cues, and the expression of appropriate effector molecules. The overall goal of this program project is to define key molecular and cellular processes that regulate effective T cell immune responses in peripheral tissue during the initial innate inflammatory response, during effector T cell activation, and during the generation and persistence of tissue-restricted memory. In concordance with this overall goal, the goal of this reagent core is to develop reagents that will enable us to both monitor and manipulate key molecules in a spatially and temporally regulated fashion in specific cell types. The reagents developed in the core will enable all of the projects to probe the functional significance of specific intracellular and membrane-associated events that change as cells interact with other cells and within specific microenvironments in inflamed tissue. The Reagent Core has three aims: 1. To generate retroviruses expressing genetic reporters to image functional events and to manipulate expression of key proteins in T cells. We will generate retroviruses that can be used to localize and manipulate structural proteins that regulate T cell migration and ECM interactions (integrins and cytoskeletal proteins), to monitor intracellular signaling events, and to modify gene expression. 2. To generate and maintain breeding stocks of genetically modified mouse strains that tag specific leukocyte populations with different fluorescent tags. To fully understand how cellular cross talk can influence an immune response, it is necessary to define the cell populations that are present within the inflamed tissue. The goal of this aim will be to maintain a breeding colony of genetically modified mouse lines that express tissue specific fluorescent reporters to identify and distinguish T cells and different myeloid cell populations in inflamed tissue. This repository will enable investigators to “mix and match” fluorescently tagged cells and recipient mice as they plan IV-MPM experiments 3. To create new mouse models to allow for identification and/or manipulation of key leukocyte subpopulations. In this aim we propose to generate several new animal models that are not currently available commercially or collaboratively. These will include mice that express leukocyte subpopulation- specific fluorescent tags, as described in Aim 2 and floxed alleles for integrins (Alpha 1 and Alpha E) that regulate the positioning and function of CD8 TRM cells.

项目总结/摘要-试剂核心 在炎症组织中，白细胞向感染部位的募集和定位是一种动态和受调节的过程。 这一过程取决于不同细胞群之间的相互作用，对特定环境的反应， 线索和适当的效应分子的表达。该项目的总体目标是 定义调节外周血中有效T细胞免疫应答的关键分子和细胞过程 在初始先天性炎症反应期间，在效应T细胞活化期间，以及在免疫应答期间， 组织限制记忆的产生和持续。为了实现这一总体目标， 试剂的核心是开发试剂，使我们能够监测和操纵关键分子， 在特定细胞类型中的空间和时间调节方式。核心中开发的试剂将使 所有的项目都是为了探测特定的细胞内和膜相关事件的功能意义 当细胞与其他细胞相互作用时，以及在发炎组织的特定微环境中，这些细胞会发生变化。的 Reagent Core有三个目标： 1.为了产生表达遗传报告基因的逆转录病毒，以成像功能事件并操纵 关键蛋白在T细胞中的表达。我们将产生逆转录病毒，可以用来定位和 操纵调节T细胞迁移和ECM相互作用的结构蛋白（整合素和细胞骨架蛋白）， 蛋白质），以监测细胞内信号传导事件和修饰基因表达。 2.为了产生和维持标记特定基因的转基因小鼠品系的育种种群， 具有不同荧光标记的白细胞群。为了充分理解细胞串扰是如何 影响免疫反应，有必要定义存在于细胞内的细胞群。 发炎的组织这一目标的目标将是保持一个繁殖群体的转基因小鼠品系 其表达组织特异性荧光报告物以鉴定和区分T细胞和不同骨髓细胞 炎症组织中的细胞群。这个储存库将使调查人员能够“混合和匹配”荧光标记 细胞和受体小鼠，因为他们计划IV-MPM实验 3.创建新的小鼠模型，以识别和/或操作关键白细胞 亚群在这个目标中，我们建议产生几个新的动物模型，目前还没有 可商购或合作获得。这些将包括表达白细胞亚群的小鼠- 如Aim 2中所述的特异性荧光标记和整合素的floxed等位基因（α 1和α E）， 调节CD 8 TRM细胞的定位和功能。