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: 10241367
• 负责人: JIM F Miller
• 金额: $ 30.96万
• 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/10241367
• 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.

项目摘要/摘要-试剂核心 炎症组织中白细胞在感染部位的募集和定位是一个动态和受调节的过程 这一过程依赖于不同细胞群体之间的串扰，对特定环境的反应 线索，以及适当的效应器分子的表达。该计划项目的总体目标是 确定调节外周T细胞有效免疫反应的关键分子和细胞过程 组织在最初的先天炎症反应期间、在效应器T细胞激活期间和在 组织限制性记忆的产生和持续。与这个总目标相一致的是，这个目标 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 在特定细胞类型中的空间和时间调节方式。在核心开发的试剂将使 所有这些项目都是为了探索特定的细胞内和膜相关事件的功能意义 当细胞与其他细胞和炎症组织中特定的微环境相互作用时，这种情况就会改变。这个 试剂核心有三个目标： 1.产生表达基因报告的逆转录病毒，以成像功能事件并操纵 T细胞中关键蛋白的表达。我们将产生逆转录病毒，可用于本地化和 操纵调节T细胞迁移和细胞外基质相互作用的结构蛋白(整合素和细胞骨架 蛋白质)，以监测细胞内信号事件，并改变基因表达。 2.产生和维护标记特定基因的转基因小鼠品系的繁育种群 具有不同荧光标记的白细胞群。要充分了解手机串扰是如何 为了影响免疫反应，有必要定义存在于 发炎的组织。这一目标的目标将是保持一个转基因小鼠品系的繁育群体 表达组织特异性荧光报告以识别和区分T细胞和不同髓系细胞 炎症组织中的种群。这个储存库将使调查人员能够“混合和匹配”荧光标记 细胞和受体小鼠计划进行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。为了达到这个目的，我们提出了几种新的动物模型，目前还没有 商业上或合作上可用的。这些将包括表达白细胞亚群的小鼠- 特定的荧光标记，如目标2和整合素的花等位基因(阿尔法1和阿尔法E)所述 调节CD8TRM细胞的定位和功能。