EFFECT OF ADHESION ON EOSINOPHIL FUNCTION

这个项目的目标是研究 运输过程中出现的嗜酸性粒细胞和粘附性胞外蛋白 嗜酸性粒细胞从血液流向呼吸道，并了解 相互作用与嗜酸性粒细胞表型的改变有关。我们 假设可能会形成导致哮喘的恶性循环 嗜酸性粒细胞渗入导致异常沉积 细胞外基质，进而导致更多的渗透和 嗜酸性粒细胞功能上调。这样做的具体目的是 建议是：(A)发展黏附和迁移的模式 嗜酸性粒细胞，强调细胞功能；(B)确定 相关基质蛋白在模型中的分布与正常， 非哮喘过敏和哮喘的支气管粘膜；(C)确定 整合素在特定目标A建立的模型中的作用；(D)确定 细胞因子和趋化因子在嗜酸性粒细胞-基质相互作用中的作用； 和(E)测试活化的嗜酸性粒细胞的产物是否改变沉积 肺成纤维细胞培养中细胞外基质的变化。要完成 这些目标，外周血嗜酸性粒细胞将被研究在两个-和 细胞黏附、扩散和迁移的三维系统。 产生超氧阴离子、白三烯C4和细胞因子，例如， 将评估GM-DSF、脱颗粒程度和细胞存活率 测定不同孵育条件下细胞功能的变化 黏附内皮细胞(VCAM-1)和基质蛋白(FN， 层粘连蛋白亚型)。细胞因子、趋化因子和 将在这些检测中检测抗整合素抗体，以确定关键 可能发生在组织内的相互作用。胶粘剂的表达 哮喘呼吸道的蛋白质将通过免疫细胞化学进行测定 原位杂交或逆转录-聚合酶链式反应。项目： 将对激活的嗜酸性粒细胞进行测试，以确定它们是否能够导致 基质蛋白沉积增加，如发生在呼吸道 哮喘患者。血嗜酸性粒细胞表型的变化 这些体外条件将与类似的评估进行比较 哮喘患者呼吸道嗜酸性粒细胞与生物学相关性研究 和临床相关性。这些观察结果应该会提供新的见解 嗜酸性粒细胞生物学和持续性呼吸道损伤机制的研究 哮喘。

The goal of this project is to examine the interactions between eosinophils and adhesive extracellular proteins occurring during transit of eosinophils from blood stream to airway and learn how the interactions relate to the alteration of eosinophil phenotype. We hypothesize that a vicious cycle may develop that contributes to asthma whereby eosinophilic infiltration causes deposition of abnormal extracellular matrix that in turn causes more infiltration and functional upregulation of eosinophils. The specific aims of this proposal are to: (A) develop models of adhesion and migration of eosinophils with an emphasis on cell function; (B) determine that distribution of relevant matrix proteins in the models vis-a-vis normal, non-asthmatic allergic and asthmatic bronchial mucosa; (C) determine the role of integrins in models established in specific aim A; (D) determine the role of cytokines and chemokines in eosinophil-matrix interactions; and (E) test whether products of activated eosinophils alter deposition of extracellular matrix in cultures of lung fibroblasts. To accomplish these goals, peripheral blood eosinophils will be studied in tow-and three-dimensional systems of cell adhesion, spreading, and migration. The production of superoxide anion, leukotriene C4, and cytokines, e.g., GM-DSF, extent of degranulation, and cellular survival will be assessed to determine alteration of cell function after various incubations with adhesive endothelial cell (VCAM-1) and matrix proteins (fibronectin, laminin isoforms). The effects of cytokines, chemokines, and antiintegrin antibodies will be examined in these assays to identify key interactions that could occur within the tissue. Expression of adhesive proteins in asthmatic airways will be determined by immunocytochemistry and in situ hybridization or reverse transcription-PCR. Projects of activated eosinophils will be tested for their ability to cause increased deposition of matrix proteins as occurs in airways of asthmatics. Changes in phenotype of blood eosinophils that result from these in vitro conditions will be compared to similar assessments on eosinophils from airways of patients with asthma to correlate biological and clinical relevance. These observations should provide new insights into eosinophil biology and mechanisms of persistent airway damage in asthma.