EFFECT OF ADHESION ON EOSINOPHIL FUNCTION

这个项目的目标是研究 嗜酸性粒细胞和粘附细胞外蛋白发生在运输过程中 嗜酸性粒细胞从血流到气道，并了解如何 相互作用与嗜酸性粒细胞表型的改变有关。 我们 假设恶性循环可能会导致哮喘 由此嗜酸性粒细胞浸润引起异常的 细胞外基质，这反过来又导致更多的浸润， 嗜酸性粒细胞的功能性上调。 具体目标是 建议是：（A）开发粘附和迁移模型， （B）确定， 模型中相关基质蛋白的分布与正态分布相比， 非哮喘性过敏性和哮喘性支气管粘膜;（C）测定 整合素在特定目标A中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 的细胞外基质。 完成 这些目标，外周血嗜酸性粒细胞将被研究在两个和 细胞粘附、扩散和迁移的三维系统。 超氧阴离子、白三烯C4和细胞因子的产生， 将评估GM-DSF、脱粒程度和细胞存活率 为了确定在用以下物质进行各种孵育后细胞功能的改变， 粘附内皮细胞（VCAM-1）和基质蛋白（纤连蛋白， 层粘连蛋白同种型）。 细胞因子、趋化因子和 抗整联蛋白抗体将在这些试验中进行检查，以确定关键的 可能发生在组织内的相互作用。 粘合剂的表达 哮喘气道中的蛋白质将通过免疫细胞化学来确定 以及原位杂交或逆转录-PCR。 项目 将测试活化的嗜酸性粒细胞引起 增加的基质蛋白沉积，如发生在 哮喘患者 血液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， 哮喘患者气道嗜酸性粒细胞与生物学 和临床相关性。 这些观察应该提供新的见解 嗜酸性粒细胞生物学和持续性气道损伤的机制， 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.