Clinical Core

这项建议的首要目标是确定呼吸道接触过敏的机制。 刺激启动外周血嗜酸粒细胞(EOS)附着并渗入肺内成为效应器 参与调节淋巴细胞和成纤维细胞从而促进呼吸道炎症的细胞 以及哮喘的重塑。我们认为IL-3刺激EOS有助于Th17淋巴细胞 通过EOS释放IL-1β和产生与组织重塑相关的因子而发展(项目 1)。IL-3刺激的EOS衍生的信号素7A的产生被假设为激活成纤维细胞以 生产细胞外基质(EClVI)成分(项目1)。其中一种成分Periostin可能会引导 进一步将EOS从血液重新招募到呼吸道(项目2)。最后，呼吸道ECM的产生也是 由EOS分泌的转化生长因子-β通过涉及顺式-反式肽基-脯氨基的信号机制调节 异构酶，Pin1(项目3)。外周血EOS的机制研究可以在体内得到验证 节段性过敏原挑战模型。拟议研究的意义在于，每个项目都将 有助于理解哮喘气道中的EOS生物学，潜在地识别出几个新的 可以改变肺重塑过程的药物靶点。要实现这一点并向 项目。核心A分为以下任务：具体任务1--招募和鉴定过敏 哮喘受试者参加涉及外周血液和呼吸道EOS的实验方案； 特定的TASL&lt；2-每周4天进行静脉采血，以净化患者的外周血EOS 过敏或过敏性哮喘患者；特异性TASL&lt；3-节段性支气管镜检查 哮喘患者支气管激发变应原(Ag-SBP)检测及BAL液和支气管镜检查 过敏原激发前后的活检(Bx)；特定任务4-执行基本处理和 对不属于核心B的呼吸道和血液样本进行实验室分析，包括免疫组织化学， 建立支气管内膜成纤维细胞系；和特殊任务5--保持高质量 符合HIPAA的数据库，包括受试者的人口统计、临床和生理特征， 安全数据和基本BAL分析，除了在数据检索、分析和 提交报告并为所有项目提供统计支持。 相关性(请参阅说明)： 鉴于EOS在哮喘严重程度进展中的核心重要性，这项提议将允许 更好地了解主动脉重塑的分子基础，并开始形成一种风险签名 可用于肺功能下降的早期检测。学习新招募和激活的能力 因路EOS应对分段变应原挑战，最大限度地促进了发现翻译。

The overarching goal of this proposal is to define the mechanisms by which airway exposure to allergic stimuli primes peripheral blood eosinophils (EOS) to attach and extravasate into tlie lung to become effector cells contributing to the regulation of lymphocytes and fibroblasts thus contributing to ainway inflammation and remodeling in asthma. We propose that IL-3 stimulation of EOS contributes to Th17 lymphocyte development through EOS release of IL-1 p and production of factors relevant to tissue remodeling (Project 1). IL-3-stimulated EOS-derived production of semaphorin7A is hypothesized to activate fibroblasts to produce extracellular matrix (EClVI) components (Project 1). One of these components, periostin, may direct further recruitment of EOS from the blood to the airway (Project 2). Finally, airway ECM production is also regulated by EOS-secreted TGF-pi via signaling mechanisms involving the cis-trans peptidyl-prolyl isomerase, Pin1 (Project 3). Mechanistic studies with peripheral blood EOS can be validated in vivo using the segmental allergen challenge model. The significance of the proposed research is that each project will contribute to the understanding of EOS biology in the asthmatic airway, potentially identifying several new drug targets that could modify the process of lung remodeling. To accomplish this and provide service to the projects. Core A is divided into the following Tasks: Specific Task 1 - To recruit and characterize allergic asthma subjects for participation in experimental protocols involving peripheral blood and airway EOS; Specific Tasl< 2 - To perform phlebotomy 4 days a week for the purification of peripheral blood EOS from patients with allergies or allergic asthma; Specific Tasl< 3 - To perform bronchoscopies with segmental bronchoprovocation with allergen (Ag-SBP) in subjects with asthma and obtain BAL fluid and endobronchial biopsies (Bx) before and after allergen challenge; Specific Task 4 - To perform basic processing and laboratory analysis of airway and blood samples that are not part of Core B, including immunohistochemistry, establishing fibroblasts cell lines from endobronchial Bx; and Specific Task 5 - To maintain high-quality HIPAA-compliant databases that includes subject demographic, clinical and physiologic characteristics, safety data, and basic BAL analysis, in addition to providing assistance in data retrieval, analysis, and presentation and to provide statistical support for all projects. RELEVANCE (See instructions): Given the central importance of EOS in the progression of asthma severity, this proposal will allow for a better understanding ofthe molecular basis of ainway remodeling and begin to develop a risl< signature that could be used for early detection of lung function decline. The ability to study newly recruited and activated ainway EOS in response to segmental allergen challenge maximally facilitates discovery translation.