Roles of CC chemokine activity in mast cell responses and ocular allergy

CC趋化因子活性在肥大细胞反应和眼部过敏中的作用

• 批准号: 7688427
• 负责人: Santa Jeremy Ono
• 金额: $ 38.75万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-08-01 至 2013-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7688427
• 关键词: Address; Adoptive Transfer; Affect; Allergens; Allergic; Allergic Conjunctivitis; Automobile Driving; Binding; Biological Assay; CC chemokine receptor 3; CCL7 gene; CCR1 gene; Cell Differentiation process; Cell physiology; Cells; Chemotaxis; Defect; Disease; Eotaxin; Gene Chips; Gene Expression; Gene Targeting; Genes; Genotype; Goals; Health; Health Care Costs; Hypersensitivity; IgE; In Vitro; Mediating; Mediator of activation protein; Modeling; Molecular; Mus; Pathogenesis; Pattern; Phase; Play; Population; Production; Proteins; Proteomics; Quality of life; RGS1 gene; Reaction; Role; Signal Transduction; Stimulus; Surface; Symptoms; Techniques; Testing; United States; Vimentin; beta-Chemokines; chemokine; crosslink; cytokine; improved; in vitro Assay; in vivo; interest; mast cell; new therapeutic target; novel; public health relevance; reconstitution; response

DESCRIPTION (provided by applicant): Allergies are a major health problem in the United States, reducing quality and life and resulting in significant health care costs. Ocular allergies affect as many as one in five people in the United States. Improved and novel treatments are clearly needed for allergic diseases. Mast cells play a critical role in ocular allergy, but the exact molecular mechanisms driving mast cell activation are poorly understood. Molecules that are necessary for ocular hypersensitivity reactions - and that might be suitable targets for treatments of ocular allergy - include the chemokines eotaxin-1 and MIP-11, their respective receptors CCR3 and CCR1, and molecules regulated by these signals. The goal of the study outlined below is to better define the roles of these molecules in ocular allergy. The study contains the following specific aims: 1) to examine the mast cell-priming role of eotaxin-1-CCR3 activity in a murine model of allergic conjunctivitis; 2) to explore the costimulatory role of MIP-11-CCR1 signaling in mast cell activation in vivo; 3) to identify proteins that mediate the mast cell effects of MIP-11-IgE costimulation in vivo; and 4) to identify gene products that, when upregulated, mediate the mast cell effects of MIP-11-IgE costimulation in vivo. The adoptive transfer model for assessing in vivo mast cell activity will be used as a component of all four aims. In this model, mast cells with a genotype of interest are injected into mast cell-deficient mice, which have a defective ocular allergy response. Reconstituted mast cell populations lacking critical genes will fail to restore early-phase and/or late-phase responses to ocular allergens. For the first two aims, mast cells deficient for eotaxin-1, MIP-11, CCR1 or CCR3 will be used for in vitro assays of mast cell differentiation and function, and for the in vivo adoptive transfer model. For the third aim, the roles of potential proteomic mediators of MIP-11-CCR1 activity, specifically vimentin and PI3K3, will be assessed using the in vitro mast cell assays and in vivo adoptive transfer model. For the fourth aim, the roles of transcriptionally upregulated mediators of MIP-11-CCR1 activity, specifically CCL7 and RGS1, will be assessed using the in vitro mast cell assays and in vivo adoptive transfer model. The results of these studies will provide a better understanding of the roles of chemokine signaling in mast cell function and ocular allergy, and may identify novel targets for therapeutic strategies. PUBLIC HEALTH RELEVANCE: Allergies are a major health problem in the United States, yet the molecular mechanisms contributing to allergic responses remain poorly defined. In this study, we will address the roles of mast cell chemokine signaling in allergic conjunctivitis. Our in vitro and in vivo techniques will clarify the roles of eotaxin-1-CCR3 signaling and MIP-11-CCR1 signaling in mast cells, and will identify relevant downstream mediators of Fc5RI- CCR1 co-activation that might serve as potential targets for novel therapies.

描述(由申请人提供)：在美国，过敏是一个主要的健康问题，降低了质量和生活，并导致了巨大的医疗费用。在美国，多达五分之一的人会受到眼睛过敏的影响。对于过敏性疾病，显然需要改进和创新的治疗方法。肥大细胞在眼部过敏中起关键作用，但肥大细胞激活的确切分子机制尚不清楚。眼部过敏反应所必需的分子--可能是治疗眼部过敏的合适靶点--包括趋化因子嗜酸性粒细胞趋化因子-1和MIP-11，它们各自的受体CCR3和CCR1，以及受这些信号调控的分子。下面概述的这项研究的目标是更好地确定这些分子在眼睛过敏中的作用。本研究有以下具体目的：1)在过敏性结膜炎小鼠模型中检测eoaxin-1-CCR3活性对肥大细胞的启动作用；2)探讨MIP-11-CCR1信号在体内肥大细胞激活中的共刺激作用；3)在体内寻找介导MIP-11-IgE共刺激肥大细胞作用的蛋白质；以及4)寻找在上调时介导体内MIP-11-IgE共刺激肥大细胞效应的基因产物。评估体内肥大细胞活性的采用转移模型将作为所有四个目标的组成部分。在这个模型中，将具有感兴趣的基因的肥大细胞注射到肥大细胞缺陷小鼠体内，这些小鼠的眼睛过敏反应有缺陷。缺乏关键基因的重组肥大细胞群体将无法恢复对眼部变应原的早期和/或晚期反应。对于前两个目标，缺乏嗜酸性粒细胞趋化因子-1、MIP-11、CCR1或CCR3的肥大细胞将用于肥大细胞的体外分化和功能检测，以及体内过继转移模型。对于第三个目标，将使用体外肥大细胞实验和体内过继转移模型来评估潜在的MIP-11-CCR1活性的蛋白质组介质，特别是波形蛋白和PI3K3的作用。对于第四个目标，将使用体外肥大细胞分析和体内过继转移模型来评估转录上调的MIP-11-CCR1活性调节因子的作用，特别是CCL7和RGS1。这些研究的结果将有助于更好地了解趋化因子信号在肥大细胞功能和眼部过敏中的作用，并可能为治疗策略确定新的靶点。公共卫生相关性：在美国，过敏是一个主要的健康问题，但导致过敏反应的分子机制仍然不清楚。在这项研究中，我们将探讨肥大细胞趋化因子信号在过敏性结膜炎中的作用。我们的体外和体内技术将阐明eoaxin-1-CCR3信号和MIP-11-CCR1信号在肥大细胞中的作用，并将识别Fc5RI-CCR1共激活的相关下游介质，这些介质可能成为新疗法的潜在靶点。