Cytoskeletal regulation of T cell-APC interactions

T 细胞-APC 相互作用的细胞骨架调节

• 批准号: 8513565
• 负责人: Anna Huttenlocher
• 金额: $ 40.98万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-01 至 2014-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8513565
• 关键词: Adhesives; Affect; Antigen-Presenting Cells; Autoimmune Diseases; Autoimmune Process; Autoimmunity; Binding; Biochemical; CD4 Positive T Lymphocytes; Cardiovascular Diseases; Cell Communication; Cell physiology; Chronic; Data; Development; Disease; Engineering; Experimental Autoimmune Encephalomyelitis; F-Actin; Focal Adhesion Kinase 1; Focal Adhesions; Foundations; Goals; Health; Human; Image; Immune; Immune response; Immunity; In Vitro; Inflammation; Integrins; Kinetics; Knock-out; Knockout Mice; Knowledge; Lead; Left; Life; Mediating; Molecular; Multiple Sclerosis; Pathogenesis; Pathologic; Pathway interactions; Phosphotransferases; Play; Positioning Attribute; Protein Isoforms; Proteins; RNA Interference; Reagent; Regulation; Research; Role; Signal Transduction; Site; Structure; Synapses; T cell regulation; T-Cell Activation; T-Lymphocyte; Talin; Testing; Three-Dimensional Imaging; Time; Transgenic Mice; Viral Tumor Antigens; fluorescence imaging; improved; in vivo; inorganic phosphate; insight; mouse model; novel; rho; therapeutic target

DESCRIPTION (provided by applicant): T cell interactions with antigen-presenting cells (APCs) are important both in normal host immune responses and in pathologic conditions including autoimmune disease. Despite their importance, we still have limited understanding of the mechanisms that regulate the formation and release of T cell-APC contacts, critical determinants of T cell activation. The long-term goal of the proposed research is to understand the molecular mechanisms that regulate the duration of T cell-APC contacts and how these 3D adhesive structures affect immune responses and the development of autoimmune disease. Although T cell-APC contact sites, known as the immune synapse, share many structural components with integrin-mediated focal adhesions, we have demonstrated that the adhesive mechanisms that regulate T cell-APC contact site dynamics are distinct. Despite this recent progress, the mechanisms that regulate the formation of transient versus sustained T cell-APC contacts to mediate T cell tolerance or immunity remain largely unknown. Moreover, the kinetics with which components enter and leave T cell-APC contact sites are not well defined. The hypothesis that guides our research is that key regulators of focal adhesion turnover, talin, phosphatidylinositide phosphate kinase PIPKI390 and focal adhesion kinase (FAK) differentially regulate the polarization of the integrin LFA-1 and the dynamic assembly and disassembly of T cell- APC contacts to affect T cell activation and autoimmunity. Substantial evidence suggests that stable T cell-APC interactions are necessary for T cell activation and the development of autoimmune disease. Yet, depletion of LFA-1, which mediates sustained T cell-APC interactions, can lead to worsening autoimmunity in the experimental autoimmune encephalitis (EAE) mouse model of multiple sclerosis, a CD4 T cell driven disease (Gultner, 2010). Therefore, the relationship between adhesive interactions and autoimmunity may be difficult to predict. Using knockout mice we have generated, we will determine how LFA-1 associated proteins, talin, PIPKI390 and FAK regulate the development of autoimmunity in EAE. We are now uniquely positioned to pursue the following specific aims: I. Test the role of talin in T cell-APC interactions and T cell activation. II. Determine how phosphatidylinositide phosphate kinase (PIPKI390) regulates T cell-APC interactions and the development of autoimmunity in EAE. III. Test the role of focal adhesion kinase (FAK) in T cell- APC interactions and T cell activation. The strength of this proposal lies in the use of conditional transgenic mice, use of novel inducible reagents and live 3D time-lapse fluorescent imaging to probe the pathways that regulate the dynamics of T cell-APC contact sites to affect T cell activation. The proposed studies will lead to improved understanding of the mechanisms that regulate T cell-APC interactions and T cell activation and may provide the foundation for new strategies to treat autoimmune disease.

描述（由申请人提供）：T细胞与抗原呈递细胞（APC）的相互作用在正常宿主免疫反应和包括自身免疫性疾病在内的病理状况中很重要。尽管它们的重要性，但我们仍然对调节T细胞APC接触形成和释放的机制的理解仍然有限，T细胞激活的关键决定因素。拟议研究的长期目标是了解调节T细胞APC接触持续时间的分子机制，以及这些3D粘合剂结构如何影响免疫反应和自身免疫性疾病的发展。尽管T细胞-APC接触位点（称为免疫突触）与整联蛋白介导的局灶性粘连共享许多结构成分，但我们已经证明调节T细胞-APC接触位点动力学的粘合机制是不同的。尽管最近取得了进展，但调节瞬态与持续的T细胞接触形成以介导T细胞耐受性或免疫力的机制在很大程度上尚不清楚。此外，组件输入并留下T细胞-APC触点位点的动力学尚未很好地定义。指导我们的研究的假设是，局灶性粘附离职率，塔林，磷脂酰素肌醇磷酸磷酸激酶PIPKI390和局灶性粘附激酶（FAK）差异调节整联蛋白LFA-1的极化和动态组合和自动化的动力学对T细胞的隔离。大量证据表明，稳定的T细胞APC相互作用对于T细胞激活和自身免疫性疾病的发展是必需的。然而，介导持续的T细胞APC相互作用的LFA-1的耗竭会导致自身免疫性恶化，在多发性硬化症的实验自身免疫性脑炎（EAE）小鼠模型，CD4 T细胞驱动的疾病中（Gultner，2010年）。因此，可能难以预测粘合剂相互作用与自身免疫性之间的关系。使用我们生成的基因敲除小鼠，我们将确定LFA-1相关的蛋白质，Talin，pipki390和FAK如何调节EAE自身免疫的发展。现在，我们可以独特地追求以下特定目的：I。测试塔林在T细胞-APC相互作用和T细胞激活中的作用。 ii。确定如何调节T细胞-APC相互作用和EAE自身免疫性的发展。 iii。测试焦点粘附激酶（FAK）在T细胞APC相互作用和T细胞激活中的作用。该提案的强度在于使用条件转基因小鼠，使用新型诱导试剂和活体3D延时荧光成像，以探测调节T细胞-APC接触位点动力学的途径，以影响T细胞的激活。拟议的研究将提高人们对调节T细胞相互作用和T细胞激活的机制的理解，并为治疗自身免疫性疾病的新策略提供基础。