INFLAMMATORY MODULATION OF BRONCHOMOTOR TONE

支气管运动张力的炎症调节

• 批准号: 6099633
• 负责人: ALAN Richard LEFF
• 金额: --
• 依托单位: UNIVERSITY OF CHICAGO
• 依托单位国家: 美国
• 财政年份: 1996
• 资助国家: 美国
• 起止时间: 1996-08-01 至 1998-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6099633
• 关键词: asthma; beta adrenergic receptor; bronchomotion; bronchus; cell adhesion; cell adhesion molecules; cooperative study; cytokine; eosinophil; fibronectins; flow cytometry; guinea pigs; human tissue; immunologic assay /test; immunomagnetic separation; inflammation; leukocyte activation /transformation; leukotrienes; muscle contraction; peroxidases; respiratory epithelium; respiratory muscles; secretion; selectins; superoxides; tissue /cell culture; vascular endothelium

Studies are proposed to examine mechanisms by which activated inflammatory cells convert normally responsive airways into hyperreactive tissues. Investigations are planned to examine the relationship between adhesion of peripheral blood and cultured eosinophils to intra- and extracellular matrix ligands and the effect of these reactions in upregulating secretion of bronchoactive granular proteins and eicosanoids. A central hypothesis of the first series of investigations is that adhesion of peripheral blood eosinophils to vascular endothelium causes augmented secretion upon stimulation. Using immunomagnetic separation techniques that isolate eosinophils in high purity from non-atopic humans developed in preliminary studies, experiments are proposed to determine the effect of preincubation of eosinophils with human umbilical vein endothelial cells (HUVEC) on the spontaneous and stimulated secretion of eosinophils peroxidase (EPO), leukotriene C4 (LTC4), and superoxide anion (O2-). Additional studies will l) examine the incremental effects on metabolic activation and secretion of eosinophil-adhesion to HUVEC pretreated with IL-1 or IL-4 and 2) determine the effect of blockade of eosinophil adhesion molecules LFA-1, Mac-1, VLA-4, and L-selectin on adhesion-mediated regulation of eosinophil secretory function. Additional experiments will address the role of exposure to matrix protein on upregulation of secretion of bioactive eosinophil proteins and eicasonoids. A second series of experiments will examine the functional correlates of endothelial adhesion and matrix protein interactions with eosinophils on airway responsiveness. Using preparations of cultured cord-derived eosinophils (CDB) developed in preliminary studies for this proposal, cell-cell interactions between CDE and excised human bronchi will be assessed in vitro and in a guinea pig "living explant" preparation in vivo. Studies are proposed to assess the effect of exogenous activation of eosinophils with formylated tripeptide and by adhesion to matrix proteins and vascular endothelium on isometric contraction and auxotonic narrowing of human airways. Using a specially developed superfusion preparation of human bronchus, the effect of epithelial activation and adhesion will be assessed in vitro by videomicrometry. In a final series of experiments, the regulation of eosinophil secretion of bronchoactive proteins and LTC4 by the beta-adrenoceptor and by endogenous phospholipase A will be related to inhibition of airway hyperresponsiveness caused by activated eosinophils. The proposed studies are designed to l) determine mechanisms causing eosinophil activation, 2) assess indices of cellular secretion corresponding to augmented airway smooth muscle contraction, and 3) model the corresponding inflammatory induction of airway hyperresponsiveness in vitro. Data derived from these investigation should provide a mechanistic basis for therapeutic intervention in the inflammatory hyperresponsiveness of human asthma.

建议进行研究以检查激活的机制 炎症细胞将正常反应的气道转化为 高反应组织。计划进行调查以审查 外周血粘附与培养物的关系 嗜酸性粒细胞与细胞内和细胞外基质配体以及 这些反应对上调分泌的作用 支气管活性颗粒蛋白和类二十烷酸。一个中央 第一系列研究的假设是粘附力 外周血嗜酸性粒细胞对血管内皮细胞影响的原因 刺激后分泌增加。使用免疫磁 分离高纯度嗜酸性粒细胞的分离技术 初步研究中开发的非特应性人类，实验是 建议确定嗜酸性粒细胞预孵育的效果 人脐静脉内皮细胞（HUVEC） 嗜酸性粒细胞过氧化物酶的自发和刺激分泌 (EPO)、白三烯 C4 (LTC4) 和超氧阴离子 (O2-)。 其他研究将 l) 检验对以下方面的增量影响： 代谢激活和嗜酸性粒细胞分泌-粘附到 HUVEC 用 IL-1 或 IL-4 预处理，并且 2) 确定 阻断嗜酸性粒细胞粘附分子 LFA-1、Mac-1、VLA-4 和 L-选择素对嗜酸性粒细胞分泌的粘附介导调节 功能。额外的实验将解决暴露的作用 基质蛋白对生物活性物质分泌的上调 嗜酸性粒细胞蛋白和类胡萝卜素。第二系列实验 将检查内皮粘附的功能相关性和 基质蛋白与气道上的嗜酸性粒细胞相互作用 反应能力。使用培养脐带来源的制剂 为此初步研究开发了嗜酸性粒细胞（CDB） 建议，CDE 和切除的人体之间的细胞间相互作用 支气管将在体外和豚鼠“活体”中进行评估 体内外植体”制备。建议进行研究以评估 外源性甲酰化嗜酸性粒细胞活化作用 三肽并通过粘附基质蛋白和血管 内皮细胞对等长收缩和张力不足变窄的影响 人类呼吸道。使用专门开发的灌注制剂 人支气管上皮活化和粘附的影响 将通过视频显微测量进行体外评估。在最后一个系列中 实验中，嗜酸性粒细胞分泌的调节 支气管活性蛋白和 LTC4 由 β-肾上腺素受体和 内源性磷脂酶A会与气道抑制有关 由活化的嗜酸性粒细胞引起的高反应性。拟议的 研究旨在 l) 确定引起嗜酸性粒细胞的机制 激活，2）评估相应的细胞分泌指数 增强气道平滑肌收缩，以及 3) 建模 气道高反应性的相应炎症诱导 体外。从这些调查中获得的数据应提供 炎症治疗干预的机制基础 人类哮喘的高反应性。