Endothelial Regulation of T Cell Subset Migration

T 细胞亚群迁移的内皮调节

基本信息

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
项目类别：
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

项目摘要

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.

根据它们产生的细胞因子，定义了不同的T细胞亚群，并执行独特的保护和病理作用。 T细胞与血管内皮细胞（EC）的分子相互作用（EC）调节效应T淋巴细胞到组织中的迁移。每个T细胞子集的趋化因子受体和粘附分子的特征在确定该子集如何迁移到组织并导致疾病。 20年前发现的Thi和Th2辅助T细胞子集分别分泌干扰素Y或IL-4，IL-5和IL-13。最近，已经确定了第三个主要的辅助T细胞子集，称为Th17。 Th17细胞分泌IL-17和IL-22，并为器官特异性自身免疫性疾病的病理以及针对某些微生物感染的保护反应做出了重要贡献。有 CDS“^ T细胞也可以分化为IL-17产生（TC17）细胞的大量证据。产生CD4*和CD8+（17型）T细胞迁移到炎症部位的迁移机制在很大程度上是未探索的相对于Interferon-y-y-Interferon-y-y-y-y-y-y-ty-thi和TC1（型）TC1（我们的TC1）的相互作用。与我们的TC1相互作用。整合素和EC，并且有 17型和1型细胞的迁移表型的显着差异。在此提案中，我们将研究17型T细胞的EC相互作用和迁移。实验计划包括三个具体目标。在AIM 1中，我们将确定17型与1型与EC相互作用的区别特征，这些特征有助于时间或空间子集特异性募集以及中性粒细胞炎症的关联与Thi7细胞。我们假设募集一个子集将影响随后的T细胞募集的选择性。我们还将探索是否与中性粒细胞浸润协调调节17型但不进行1型效应T细胞的募集。实验方法将包括对原代小鼠THI7和TCL7细胞中相关分子的蛋白质和mRNA表达的分析，以及与T细胞相互作用与与T细胞相互作用的研究重组粘附分子和在生理流动条件下的体外内皮。 AIM 2将重点放在获得17型细胞的选择素结合表型所需的分子信号上，并且将与1型T型T细胞中所需的不同信号进行比较。实验方法将涉及与选择蛋白配体表达相关的不同细胞因子信号和下游转录因子的分析，并特别关注Roryt和TBET的对比作用，Roryt和TBET是Th17和Th1细胞的标志性转录调节剂。 AIM 3将比较体内17型和1型效应T细胞的募集。实验方法将包括转移将THI7，TCL7，TH1和TC1种群明显标记为不同的小鼠炎症模型。宿主小鼠中17型和1型T细胞的迁移行为将通过几种定量技术进行评估。在AIM 1中鉴定的粘附途径和趋化因子将在这些模型中使用抗体阻断和T细胞或宿主小鼠在粘附分子和趋化因子上缺乏的宿主小鼠进行评估。受体。与项目1和3的协作研究将重点关注Th17-和TC17-内皮相互作用的CD47和整合素依赖性机制。从这些研究中获得的信息对于充分了解T细胞介导的疾病的重要募集阶段至关重要，数据将有助于确定特定于T细胞子集的治疗靶标。