Endothelial Regulation of T Cell Subset Migration

T 细胞亚群迁移的内皮调节

• 批准号: 7753044
• 负责人: ANDREW H LICHTMAN
• 金额: $ 48.95万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-07-01 至 2014-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7753044
• 关键词: AKAP9 gene; Address; Adhesions; Adhesives; Affect; Allografting; Antibodies; Antigens; Autoimmune Diseases; Autoimmune Process; Automobile Driving; Behavior; Binding; Blocking Antibodies; Brain; CD4 Positive T Lymphocytes; CD47 gene; CD8B1 gene; Cell Adhesion; Cell Adhesion Molecules; Cell Communication; Cells; Characteristics; Complex; Cytokine Signaling; Data; Disease; Disease model; E-Selectin; Endothelial Cells; Endothelium; Functional disorder; Genes; Goals; Heart; Helper-Inducer T-Lymphocyte; Homing; Immune System Diseases; In Vitro; Infection; Infiltration; Inflammation; Inflammatory; Integrins; Interferons; Interleukin-13; Interleukin-17; Interleukin-4; Interleukin-5; Interleukin-6; Intestines; Islets of Langerhans; Knock-out; Knockout Mice; Knowledge; Label; Learning; Life; Ligands; Link; Mediating; Methods; Microbe; Modeling; Molecular; Mus; Organ; P-Selectin; Pathologic; Pathology; Pathway interactions; Peritoneum; Phase; Phenotype; Physiological; Play; Population; Principal Investigator; Process; Proteins; Publishing; Recombinants; Recruitment Activity; Regulation; Relative (related person); Research Design; Role; STAT3 gene; Selectins; Signal Transduction; Site; Skin; T-Lymphocyte; T-Lymphocyte Subsets; Techniques; Testing; Th1 Cells; Th2 Cells; Tissues; Transduction Gene; Transgenic Mice; Transgenic Organisms; Vascular Endothelial Cell; Viral; Work; base; cell motility; cell type; chemokine; chemokine receptor; cytokine; in vivo; interleukin-22; interleukin-23; intravital microscopy; mRNA Expression; microbial; migration; neutrophil; programs; research study; response; small hairpin RNA; therapeutic target; transcription factor

Different T cell subsets are defined on the basis of the cytokines they produce, and they perform distinct protective and pathologic roles. Migration of effector T lymphocytes into tissues is regulated by molecular interactions of the T cells with vascular endothelial cells (EC). The chemokine receptors and adhesion molecules characteristic of each T cell subset play essential roles in determining how that subset migrates into tissues and contributes to disease. Thi and Th2 helper T cell subsets, discovered over 20 years ago, secrete interferon y or IL-4, IL-5 and IL-13, respectively. Recently, a third major helper T cell subset called Th17 has been identified. Th17 cells secrete IL-17 and IL-22, and contribute in significant ways to the pathology of organ specific autoimmune diseases, as well as to protective responses against certain microbial infections. There is substantial evidence that CDS"^ T cells can differentiate into IL-17 producing (Tc17) cells as well. The mechanisms of migration of IL-17 producing CD4* and CD8+ (Type 17) T cells into inflammatory sites are largely unexplored relative to interferon-y producing Thi and Tc1 (Type 1) T cells. Our preliminary data indicate that Th17 cells engage in adhesive interactions with selectins, integrins, and EC, and that there are significant differences in the migratory phenotypes of Type 17 and Type 1 cells. In this proposal, we will study EC interactions and migration of Type 17 T cells. The experimental program includes three Specific Aims. In Aim 1, we will determine the distinguishing features of Type 17 vs. Type 1 interactions with EC, which contribute to temporal or spatial subset specific recruitment and to the association of neutrophilic inflammation with Thi7 cells. We hypothesize that recruitment of one subset will impact on the selectivity of subsequent T cell recruitment. We will also explore if Type 17, but not Type 1 effector T cell recruitment is coordinately regulated with neutrophilic infiltration. The experimental approach will include analyses of protein and mRNA expression of relevant molecules in primary mouse Thi7 and Tcl7 cells and studies of T cell interactions with recombinant adhesion molecules and with endothelium in vitro under physiological flow conditions. Aim 2 will focus on the molecular signals needed for the acquisition of the selectin-binding phenotype of Type 17 cells, and comparisons will be made with the different signals required selectin ligand expression in Type 1 T cells. The experimental approach will involve analysis of different cytokine signals and downstream transcription factors linked to expression of selectin ligands, with a particular focus on the contrasting roles of RORyT and Tbet, which are the signature transcriptional regulators of Th17 and Th1 cells. Aim 3 will compare the recruitment of Type 17 and Type 1 effector T cells in vivo. The experimental approach will include transfer of distinctly labeled Thi7, Tcl7, Th1 and Tc1 populations into different mouse inflammatory models. The migratory behavior of the Type 17 and Type 1 T cells in the host mice will be assessed by several quantitative techniques. The adhesion pathways and chemokines identified in Aim 1 will be evaluated in these models using antibody blocking and T cells or host mice genetically deficient in adhesion molecules and chemokine receptors. Collaborative studies with Projects 1 and 3 will focus on CD47- and integrin-dependent mechanisms of Th17- and Tc17-endothelial interactions. The information obtained from these studies will be essential for a full understanding the important recruitment phase of T cell-mediated diseases, and the data will help determine therapeutic targets that are T cell subset specific.

不同的T细胞亚群是根据它们产生的细胞因子来定义的，它们发挥着不同的保护和病理作用。T细胞与血管内皮细胞(EC)之间的分子相互作用调节着效应T细胞向组织内的迁移。每个T细胞亚群特有的趋化因子受体和黏附分子在决定该亚群如何迁移到 组织和致病因素。20多年前发现的Th1和Th2辅助T细胞亚群分别分泌干扰素y或IL-4、IL-5和IL-13。最近，第三个主要的辅助性T细胞亚群Th17已经被发现。Th17细胞分泌IL-17和IL-22，在器官特异性自身免疫性疾病的病理以及对某些微生物感染的保护性反应中起着重要作用。的确有 大量证据表明CDS“^T细胞也可以分化为产生IL-17的(TC17)细胞。相对于产生干扰素的THI和Tc1(类型1)T细胞，产生IL-17的CD4*和CD8+(17型)T细胞向炎症部位迁移的机制在很大程度上是未知的。我们的初步数据表明，Th17细胞与选择素、整合素和EC进行黏附相互作用，并且存在 17型和1型细胞的迁移表型有显著差异。在这项提案中，我们将研究17型T细胞的EC相互作用和迁移。该实验计划包括三个具体目标。在目标1中，我们将确定与EC的17型和1型相互作用的区别特征，这有助于时间或空间亚集的特异性招募和中性粒细胞炎症的关联 使用Thi7细胞。我们假设，一个亚群的招募将影响随后T细胞招募的选择性。我们还将探索17型而不是1型效应器T细胞募集是否与中性粒细胞渗透协调调节。实验方法将包括分析相关分子在原代小鼠Thi7和Tcl7细胞中的蛋白质和mRNA表达，以及研究T细胞与 重组黏附分子与血管内皮细胞在体外生理流动条件下。目的2将重点介绍获得17型细胞选择素结合表型所需的分子信号，并与1型T细胞表达选择素配体所需的不同信号进行比较。 实验方法将涉及分析与选择素配体表达有关的不同细胞因子信号和下游转录因子，特别关注作为Th17和Th1细胞标志性转录调节因子的RORyT和Tbet的对比作用。目的3将比较17型和1型效应T细胞在体内的募集情况。试验性方法将包括将 明确标记Thi7、Tcl7、Th1和Tc1群体进入不同的小鼠炎症模型。17型和1型T细胞在宿主小鼠体内的迁移行为将通过几种定量技术进行评估。在AIM 1中确定的黏附途径和趋化因子将在这些模型中使用抗体阻断和T细胞或缺乏黏附分子和趋化因子的遗传缺陷的宿主小鼠进行评估 感受器。与项目1和项目3的合作研究将集中在CD47和整合素依赖的Th17和Tc17-内皮相互作用的机制上。从这些研究中获得的信息对于全面了解T细胞介导的疾病的重要招募阶段是必不可少的，这些数据将有助于 确定T细胞亚群特异性的治疗靶点。