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.

项目摘要/摘要 – 试剂核心 白细胞募集和定位到发炎组织的感染部位是一种动态且受调节的过程 该过程取决于不同细胞群之间的串扰、对特定环境的反应 线索以及适当效应分子的表达。该计划项目的总体目标是 定义调节外周有效 T 细胞免疫反应的关键分子和细胞过程 在初始先天性炎症反应期间、效应 T 细胞激活期间和 组织限制记忆的产生和持续。与这一总体目标相一致，本次的目标 试剂的核心是开发试剂，使我们能够监测和操纵关键分子 特定细胞类型中空间和时间调节的时尚。核心开发的试剂将使 所有旨在探索特定细胞内和膜相关事件的功能意义的项目 当细胞与其他细胞相互作用以及发炎组织的特定微环境内时，这种变化就会发生变化。这 Reagent Core 具有三个目标： 1. 生成表达基因报告基因的逆转录病毒，以成像功能事件并操纵 T 细胞中关键蛋白的表达。我们将生成可用于本地化和 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 蛋白质），监测细胞内信号事件，并修改基因表达。 2. 产生并维持带有特定标签的转基因小鼠品系的种畜 具有不同荧光标签的白细胞群。充分了解细胞串扰如何 影响免疫反应，有必要定义存在于免疫反应中的细胞群 发炎的组织。该目标的目标是维持转基因小鼠品系的繁殖群体 表达组织特异性荧光报告基因以识别和区分 T 细胞和不同的骨髓细胞 发炎组织中的群体。该存储库将使研究人员能够“混合搭配”荧光标记 细胞和受体小鼠计划 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 亚人群。为了这个目标，我们建议产生几种目前还没有的新动物模型 可商业或合作获得。这些将包括表达白细胞亚群的小鼠 - 特定荧光标签，如目标 2 中所述，以及整联蛋白的 floxed 等位基因（Alpha 1 和 Alpha E） 调节 CD8 TRM 细胞的定位和功能。