T cell function in asthma depends on a novel signaling pathway

哮喘中的 T 细胞功能依赖于一种新的信号通路

基本信息

批准号：
7373462
负责人：
JULIA K L WALKER
金额：
$ 39万
依托单位：
DUKE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2008
资助国家：
美国
起止时间：
2008-02-01 至 2013-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7373462
关键词：
Adam11 gene Address Adrenal Cortex Hormones Agonist Allergens Allergic Anatomy Animal Model Antigens Arrestin Arrestins Asthma Binding CCL22 gene CD4 Positive T Lymphocytes CXCL10 gene CXCR3 gene Cell Adhesion Molecules Cell physiology Cells Chemotaxis Chronic Co-Immunoprecipitations Complex Data Development Development, Other Disease Dyes Eosinophilia Event Extrinsic asthma GTP-Binding Proteins Generations Goals Harvest Human Immune In Vitro Inflammatory Inflammatory Response Investigation Label Left Leukotrienes Location Lung Lymphoid Tissue Mediating Mediator of activation protein Methods Modeling Mus Ovalbumin Pathway interactions Pharmacotherapy Receptor Cell Receptor Signaling Regulation Research Role Route Second Messenger Systems Severities Signal Pathway Signal Transduction Symptoms T-Cell Activation T-Cell Receptor T-Lymphocyte Testing Th1 Cells Th2 Cells Therapeutic Transmembrane Domain Wild Type Mouse Work airway hyperresponsiveness allergic airway disease arrestin 2 base beta-Chemokines chemokine receptor chlorambucil/dactinomycin/methotrexate protocol concept desensitization immune function in vitro Assay in vivo inhibitor/antagonist member migration novel pathogen prevent receptor research study response scaffold second messenger

项目摘要

DESCRIPTION (provided by applicant): The primary goal of this project is to delineate the mechanism by which arrestin-2 regulates T cell chemotaxis in asthma. We previously showed that mice devoid of arrestin-2 do not develop allergic airway disease (AAD) and that T cell chemotaxis is impaired in these mice. Our preliminary data extend these observations to specifically show that Th2 cell chemotaxis is impaired in the absence of arrestin-2. Furthermore, we propose that the mechanism by which arrestin-2 positively regulates Th2 cell chemotaxis is through use of the novel arrestin-2-dependent signaling pathway. Our preliminary data also suggest that Th1 cell chemotaxis may be positively regulated by arrestin-2. Although not well understood, Th1 cells also contribute to the development of allergic inflammatory airway disease and thus, their regulation by arrestin-2 will be considered in this project. The schema to be addressed is: Agonist activation of chemokine receptor results in the assembly of a chemokine receptor/arrestin-2/MAPK signaling complex, which induces antigen-polarized T cell chemotaxis. Primary T cells that transgenically express an OVA-specific T cell receptor (TgTCR), replete with, and devoid of, arrestin-2, will be Th1- or Th2-polarized both in vitro and in vivo to test the hypotheses listed in the following Specific Aims: 1. To demonstrate that agonist binding of CCR4 leads to activation of a arrestin-2-dependent cell signaling mechanism in Th2 cells. 2. To demonstrate that the arrestin-2-dependent signaling pathway significantly promotes chemotaxis of allergen-activated [Th1 and Th2] cells. 3. To demonstrate that chemotaxis of [Th1 and Th2] cells from secondary lymphoid tissue to the lung is impaired in cells devoid of arrestin-2. We will use an in vivo competition transfer model where CD4+ T cells expressing an OVA-specific TCR are harvested from WT-TgTCR and arrestin-2-KO-TgTCR mice, fluorescently labeled with a stable intracellular dye, and transferred to a recipient mouse. Results from these aims will define key migratory routes for Th1 and Th2 cells in AAD, will define an arrestin-dependent chemokine receptor signaling pathway and will integrate these themes to show that arrestin-dependent signaling underlies the development of AAD. New concepts emanating from this research may provide new strategies for the treatment for asthma whereby therapeutics are developed that can selectively regulate arrestin-dependent signaling pathways, while leaving G protein-dependent signaling pathways untouched. Project Narrative: Asthma cannot be established unless T cells, a type of inflammatory cell, are able to migrate to the lung. Although detrimental in asthma, T cells are very important for immune defense against pathogens. Our work will provide information that could revolutionize asthma treatment by aiding the search for a therapeutic that can selectively prevent the deleterious migration of T cells while leaving their beneficial effects untouched.

描述（由申请人提供）：该项目的主要目标是描述抑制蛋白-2 调节哮喘中 T 细胞趋化性的机制。我们之前表明，缺乏抑制蛋白-2 的小鼠不会患上过敏性气道疾病 (AAD)，并且这些小鼠的 T 细胞趋化性受损。我们的初步数据扩展了这些观察结果，具体表明，在缺乏抑制蛋白-2 的情况下，Th2 细胞趋化性受到损害。此外，我们提出，arrestin-2 正向调节 Th2 细胞趋化性的机制是通过使用新的抑制蛋白-2 依赖性信号通路。我们的初步数据还表明 Th1 细胞趋化性可能受到抑制蛋白-2 的正向调节。尽管尚未充分了解，但 Th1 细胞也会导致过敏性炎症性气道疾病的发生，因此本项目将考虑抑制蛋白-2 对它们的调节。要解决的方案是：趋化因子受体的激动剂激活导致趋化因子受体/arrestin-2/MAPK信号复合物的组装，从而诱导抗原极化的T细胞趋化性。转基因表达 OVA 特异性 T 细胞受体 (TgTCR) 的原代 T 细胞，充满或不含抑制蛋白 2，将在体外和体内进行 Th1 或 Th2 极化，以测试以下列出的假设具体目标： 1. 证明 CCR4 的激动剂结合导致 Th2 细胞中抑制蛋白 2 依赖性细胞信号传导机制的激活。 2. 证明视紫红质抑制蛋白2依赖性信号通路显着促进过敏原激活的[Th1和Th2]细胞的趋化性。 3. 证明在缺乏抑制蛋白-2 的细胞中，[Th1 和 Th2] 细胞从次级淋巴组织到肺部的趋化性受到损害。我们将使用体内竞争转移模型，其中表达 OVA 特异性 TCR 的 CD4+ T 细胞从 WT-TgTCR 和arrestin-2-KO-TgTCR 小鼠中收获，用稳定的细胞内染料进行荧光标记，然后转移到受体小鼠中。这些目标的结果将定义 AAD 中 Th1 和 Th2 细胞的关键迁移途径，定义抑制蛋白依赖性趋化因子受体信号通路，并将整合这些主题以表明抑制蛋白依赖性信号传导是 AAD 发展的基础。这项研究产生的新概念可能为哮喘治疗提供新策略，从而开发出可以选择性调节视紫红质抑制蛋白依赖性信号通路，同时不影响 G 蛋白依赖性信号通路的治疗方法。项目叙述：除非 T 细胞（一种炎症细胞）能够迁移到肺部，否则无法确定哮喘。尽管 T 细胞对哮喘有害，但它对于抵抗病原体的免疫防御非常重要。我们的工作将提供信息，通过帮助寻找一种治疗方法，可以彻底改变哮喘治疗，这种治疗方法可以选择性地阻止 T 细胞的有害迁移，同时不影响其有益作用。