Stability and Plasticity of Regulatory T Cells

调节性 T 细胞的稳定性和可塑性

• 批准号: 7688823
• 负责人: Abul K. Abbas
• 金额: $ 19.73万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-05-01 至 2014-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7688823
• 关键词: Address; Allografting; Antigens; Autoantigens; Autoimmune Diseases; Autoimmune Process; Autoimmunity; Automobile Driving; Biochemical; Biological; Biological Assay; Breeding; CD4 Positive T Lymphocytes; Cell Differentiation process; Cell Lineage; Cell Separation; Cell surface; Cells; Chimeric Proteins; Clinical; Complement; Development; Disease; Effector Cell; Environment; Eragrostis; Exposure to; Figs - dietary; Flow Cytometry; Foundations; Gene Expression; Generations; Genes; Genetic; Goals; Harvest; Homeostasis; Human; IL2RA gene; Immune; Immunotherapy; In Vitro; Inbred NOD Mice; Inflammation; Inflammatory; Insulin-Dependent Diabetes Mellitus; Interleukin-10; Interleukin-17; Knockout Mice; Knowledge; Memory; Methods; Methylation; Molecular; Molecular Profiling; Mouse Strains; Mus; Neuropilin-1; Normal Cell; Ovalbumin; Pathway interactions; Patients; Peripheral; Peripheral Blood Mononuclear Cell; Phase; Phenotype; Philosophy; Play; Population; Production; Reaction; Recording of previous events; Reporter; Role; Signal Pathway; Signal Transduction; Sorting - Cell Movement; Specificity; Staining method; Stains; Stimulus; Surface; System; T-Lymphocyte; T-Lymphocyte Subsets; Technology; Testing; Therapeutic; Thymus Gland; Time; Transfection; Transgenic Mice; Transgenic Organisms; Translating; Tretinoin; autoreactivity; base; clinical phenotype; cytokine; design; flu; graft vs host disease; healthy volunteer; in vivo; islet; knock-down; molecular marker; mouse model; mutant; novel; novel strategies; peripheral blood; prevent; programs; promoter; research study; response; small hairpin RNA; success; transcription factor

The objective of this project is to define the plasticity and stability of regulatory T cells (Tregs) and the possibility of converting effector cells to functional Tregs, as a prelude to cellular therapy. Our hypothesis is that activated T cells go through a phase of transient Foxp3 expression that is a critical decision point, and stable FoxpS expression is a key determinant of Treg stability. Our approach is to exploit mouse models to address these questions in vivo and in vitro and to develop predictive phenotypic and molecular markers for Treg stability, and use this information to define the stability of human Tregs and to genetically manipulate human T cells in order to program their differentiation pathway. The following Specific Aims are proposed. 1. Functional and phenotypic plasticity of regulatory T cells (Tregs) and effector T cells (TefO. In this Aim, we will generate antigen-specific natural (thymic) and adaptive (peripheral) Tregs using TCR transgenic mice expressing fluorescent reporters for FoxpS and key cytokines (IFN-v, IL-17 and IL-10) and different forms of antigen exposure. Purified Tregs will be transferred into normal recipients and exposed to antigen with and without inflammatory stimuli, or transferred into recipients with active autoimmune reactions. The cells will be followed for FoxpS and cytokine expression and surface phenotype by staining, and unctional responses after sorting. Tregs will be isolated from healthy volunteers and patients with autoimmune disease (initially T1D) and followed in vitro for conversion to effectors in the presence of activating and inflammatory stimuli. A novel transgenic mouse strain that expresses a lineage marker of past roxp3 expression will be used to examine the fates of these cells. The conversion of effector cells into Tregs be examined using the same approaches. 2.'Biochemical basis of Treg stability and conversion. T cells will be examined under conditions of stability and conversion for changes in surface phenotype, expression of Treg-specific genes, and methylation of the :oxp3 locus. The functional consequence of transient FoxpS expression will be analyzed in mouse and human cells using shRNA knockdowns and knockout mice. Attempts will be made to maintain the stability of regs by expressing selected genes in human Tregs.

本项目的目的是确定调节性T细胞（Tcells）的可塑性和稳定性，以及Tcells的表达。 将效应细胞转化为功能性T细胞的可能性，作为细胞治疗的前奏。我们的假设是 活化的T细胞经历瞬时Foxp 3表达的阶段，这是一个关键的决定点， 稳定的FoxpS表达是Treg稳定性的关键决定因素。我们的方法是利用小鼠模型， 在体内和体外解决这些问题，并开发预测表型和分子标记， Treg稳定性，并使用该信息来定义人Treg的稳定性，并进行基因操作。 人类T细胞，以编程其分化途径。提出了以下具体目标。 1.调节性T细胞（TcR）和效应T细胞（TcO）的功能和表型可塑性。 在这个目标中，我们将使用TCR产生抗原特异性的天然（胸腺）和适应性（外周）T细胞。 表达FoxpS和关键细胞因子（IFN-γ、IL-17和IL-10）的荧光报告基因的转基因小鼠， 不同形式的抗原暴露。将纯化的甲状腺素转移到正常受体中，并暴露于 抗原与和没有炎症刺激，或转移到受体与积极的自身免疫反应。 通过染色跟踪细胞的FoxpS和细胞因子表达以及表面表型， 分类后的功能反应。将从健康志愿者和患有 自身免疫性疾病（最初为T1 D），然后在体外在存在 激活和炎症刺激。一种新的转基因小鼠品系， roxp 3的表达将被用于检查这些细胞的命运。效应细胞转化为T细胞 使用相同的方法进行检查。 2. Treg稳定性和转化的生物化学基础。将在稳定条件下检查T细胞 以及表面表型变化、Treg特异性基因表达和Treg基因甲基化的转化。 ：oxp 3基因座。瞬时FoxpS表达的功能性后果将在小鼠中进行分析， 使用shRNA敲除和敲除小鼠的人类细胞。将努力保持稳定， 通过在人THP中表达选定的基因来表达。