CELLULAR REGULATION OF PULMONARY INFLAMMATION AND HOST DEFENSE

肺部炎症和宿主防御的细胞调节

• 批准号: 6564870
• 负责人: Christopher B. Wilson
• 金额: $ 28.24万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-12-01 至 2002-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6564870
• 关键词: adult respiratory distress syndrome; alveolar macrophages; apoptosis; bacterial disease; cell adhesion molecules; cellular pathology; cytokine; genetically modified animals; immunopathology; inflammation; laboratory mouse; lipopolysaccharides; lung injury; nuclear factor kappa beta; pulmonary surfactants; respiratory epithelium; respiratory infections; vascular endothelium permeability

Complex processes govern lung inflammation and defenses against infection. ARDS appears to represent a disordered lung inflammatory response. The development of new therapies for this disorder are likely only to follow improved understanding of the fundamental processes controlling lung inflammation. This project seeks to explore the role in lung inflammation of two cell types poised at the anatomical interface between the lung and external environment: airway and alveolar epithelial cells and lung macrophages, and in these cells of NF-kappa B. NF-kappaB proteins play a key role in the transcriptional regulation of multiple genes, the products of which regulate inflammation, immunity and apoptosis. This project will address the following specific aims: (1) To determine the role of NF-kappa B dependent gene expression by the distal airway and alveolar epithelium in the lung response to LPS or bacterial infection, through the generation of transgenic mice in which dominant negative IkBa or RelB is expressed selectively in these cells under the control of the surfactant apoprotein C promoter. (2) To determine whether varying the time and magnitude of NF- kappaB modulation in distal airway and alveolar epithelium alters the nature of the lung response to LPS or bacterial infection. This will be done by generating transgenic mice in which expression of the ecdysone receptor is directed to these cell types under the control of the surfactant apoprotein C promoter and dominant negative IkB is expressed under the control of the surfactant apoprotein C promoter and dominant negative IkB is expressed under the control of an ecdysone responsive promoter. (3) To compare the role of NF-kappaB dependent gene transcription by lung macrophages versus lung epithelial cells in the pulmonary response to LPS or bacterial infection. This aim will be addressed by comparing the response in transgenic mice in which dominant negative IkBa is expressed selectively in macrophages under the control of the CD68 or lysosome gene regulatory elements, to the response of mice described in Aims 1 and 2. For comparison liposome-encapsulated clordronate mediated depletion of alveolar macrophages will be used, since this is an established method to assess the contribution of this cell population to lung inflammation and infection. The response to LPS and bacterial (K pneumoniae and P aeruginosa) challenges to the lung will be assessed in the different groups of mice through assessment of neutrophil recruitment, lung vascular leak, cytokine/chemokine and adhesion molecular expression, and alterations in lung surfactant aproprotein expression and apoptosis. Together these aims will evaluate, using novel reagents and approaches, the role of lung epithelium and lung macrophages in the regulation of lung inflammation, defense and cellular homeostasis.

复杂的过程控制肺部炎症和抵抗感染。 ARDS似乎代表了一种紊乱的肺部炎症反应。的 这种疾病的新疗法的发展可能只会遵循 提高对控制肺的基本过程的理解 炎症该项目旨在探索在肺部炎症中的作用 两种细胞类型的平衡在肺和肺之间的解剖界面， 外部环境：气道和肺泡上皮细胞和肺 巨噬细胞，并在这些细胞的NF-κ B B。NF-kappaB蛋白在肿瘤的发生发展中起着重要的作用。 在多个基因的转录调控中起关键作用， 其调节炎症、免疫和细胞凋亡。该项目将 具体目的如下：（1）确定NF-κ B的作用 远端气道和肺泡上皮细胞的B依赖性基因表达 在肺部对LPS或细菌感染的反应中，通过产生 表达显性阴性IkBa或RelB的转基因小鼠 在表面活性剂脱辅基蛋白的控制下， C启动子。(2)为了确定是否改变NF的时间和幅度- 远端气道和肺泡上皮中的kappaB调节改变了 肺对LPS或细菌感染反应的性质。这将是 通过产生转基因小鼠来完成， 受体是针对这些细胞类型的控制下， 表面活性剂载脂蛋白C启动子和显性负IkB表达 在表面活性剂载脂蛋白C启动子的控制下， 负性IkB在蜕皮激素应答的控制下表达， 启动子(3)比较NF-kappaB依赖基因在肿瘤细胞中的作用 肺巨噬细胞与肺上皮细胞的转录 肺对LPS或细菌感染的反应。这一目标将是 通过比较转基因小鼠的反应来解决， 阴性IkBa在巨噬细胞中选择性表达， CD 68或溶酶体基因调控元件对小鼠的反应 目标1和2中所述。为了比较，脂质体包封 将使用氯膦酸盐介导的肺泡巨噬细胞耗竭，因为 这是一种评估该细胞贡献的既定方法 人群肺部炎症和感染。对LPS的反应， 细菌（肺炎克雷伯菌和铜绿假单胞菌）对肺部的挑战将是 在不同组小鼠中通过评估中性粒细胞 募集、肺血管渗漏、细胞因子/趋化因子和粘附分子 表达和肺表面活性物质前体蛋白表达的改变， 凋亡这些目标将一起评估，使用新的试剂和 方法，肺上皮细胞和肺巨噬细胞的作用， 调节肺部炎症、防御和细胞稳态。