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，并且它们执行不同的保护和病理作用。效应T淋巴细胞向组织中的迁移受T细胞与血管内皮细胞（EC）的分子相互作用的调节。每个T细胞亚群的趋化因子受体和粘附分子特征在决定该亚群如何迁移到 组织并导致疾病。20多年前发现的Th 1和Th 2辅助性T细胞亚群分别分泌干扰素γ或IL-4、IL-5和IL-13。最近，第三个主要的辅助性T细胞亚群被称为Th 17已被确定。Th 17细胞分泌IL-17和IL-22，并以重要方式促进器官特异性自身免疫性疾病的病理学，以及对某些微生物感染的保护性应答。有 CD 17 + T细胞也可以分化成IL-17产生细胞（Tc 17）的大量证据。相对于产生干扰素-γ的Thi和Tc 1（1型）T细胞，产生IL-17的CD 4 * 和CD 8+（17型）T细胞迁移到炎症部位的机制在很大程度上尚未探索。我们的初步数据表明，Th 17细胞与选择素、整合素和EC进行粘附相互作用， 17型和1型细胞迁移表型的显著差异。在本提案中，我们将研究EC相互作用和17型T细胞的迁移。实验计划包括三个具体目标。在目标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亚群特异性募集以及与嗜酸性炎症的相关性。 Th 7细胞我们假设一个亚群的募集将影响后续T细胞募集的选择性。我们还将探索17型效应T细胞募集是否与嗜中性粒细胞浸润协调调节，而不是1型效应T细胞募集。实验方法将包括分析原代小鼠Thi 7和Tcl 7细胞中相关分子的蛋白质和mRNA表达，以及研究T细胞与Thi 7细胞的相互作用。 重组粘附分子和生理流动条件下的体外内皮细胞。目标2将集中在17型细胞的选择素结合表型的收购所需的分子信号，并进行比较与1型T细胞中的选择素配体表达所需的不同信号。 实验方法将涉及不同的细胞因子信号和下游转录因子的分析与选择素配体的表达，特别关注RORyT和Tbet的对比作用，这是Th 17和Th 1细胞的签名转录调节因子。目的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 在不同的小鼠炎症模型中观察到不同标记的Th 7、Tcl 7、Th 1和Tc 1群体。将通过几种定量技术评估宿主小鼠中17型和1型T细胞的迁移行为。将在这些模型中使用抗体阻断和T细胞或粘附分子和趋化因子遗传缺陷的宿主小鼠评价Aim 1中鉴定的粘附途径和趋化因子 受体。与项目1和3的合作研究将集中在Th 17和Tc 17-内皮相互作用的CD 47和整合素依赖性机制。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募集阶段至关重要，这些数据将有助于 确定T细胞亚群特异性的治疗靶点。