PROTEASES AND ANTIPROTEASES IN LUNG: EFFECTS OF SMOKING

肺中的蛋白酶和抗蛋白酶：吸烟的影响

• 批准号: 3485395
• 负责人: AARON JANOFF
• 金额: $ 22.04万
• 依托单位: STATE UNIVERSITY NEW YORK STONY BROOK
• 依托单位国家: 美国
• 财政年份: 1976
• 资助国家: 美国
• 起止时间: 1976-12-01 至 1991-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3485395
• 关键词: adult respiratory distress syndrome; affinity chromatography; alveolar macrophages; antibody specificity; antioxidants; bronchoscopy; disease /disorder model; elastases; electron microscopy; emphysema; enzyme linked immunosorbent assay; enzyme structure; histamine release; human subject; human tissue; immunoperoxidase; leukocytes; lung; mast cell; methionine; oxidation; peptidases; protease inhibitor; proteoglycan; psychobiology; pulmonary surfactants; respiratory function; respiratory pharmacology; tissue /cell culture

There are five specific aims: 1) We will attempt to detect and localize oxidized alpha 1-proteinase inhibitor in lung biopsies obtained from smoking and sham-smoking baboons. For this purpose, a specific monoclonal antibody that recognizes only the oxidized inhibitor will be used as the primary staining reagent, followed by established secondary-antibody immunohistochemical methods (peroxidase-antiperoxidase, avidin-biotin). This objective is important to human health because oxidation of alpha 1- proteinase inhibitor in the lungs by cigarette smoking has been implicated as a major pathogenetic factor in pulmonary emphysema, but remains unproven. 2) We will explore the role of a newly discovered mast cell elastase in the pathogenesis of smokers' emphysema in three ways: (a) by monitoring release of the enzyme (using ELISA-techniques) from purified human lung mast cells incubated with aqueous solutions of tobacco smoke and smoke fractions; (b) by measuring levels of histamine (a marker of mast cell secretory response) in bronchoalveolar lavage fluids of human smokers (before and after smoking) and of rats following acute exposure to inhaled cigarette smoke; and (c) by attempting to induce experimental emphysema in rats and dogs using repeated intrapulmonary instillations of mast cell secretogogues. 3) We will attempt to develop new inhibitors of human neutrophil elastase (a protease suspected on being a pathogenetic factor in pulmonary emphysema) based on our recent characterization of a specific neutrophil elastase inhibitor in pneumococci. 4) We will test an oxidation-resistant, genetically-engineered, mutant (Met 358Val) alpha 1-proteinase inhibitor for its resistance to inactivation in vivo in an established animal model of acute cigarette smoke exposure. 5) Finally, we will measure levels o f the neutrophil-monocyte marker antigen (L1) in blood and airspace secretions of septic and traumatized patients at risk for adult respiratory distress syndrome (ARDS) and in those with frank ARDS. Levels of the L1 protein may prove to have predictive value of ARDS in high-risk subjects.

有五个具体目标：1）我们将尝试检测和 在肺活检中定位氧化型 α1-蛋白酶抑制剂 从吸烟和假吸烟的狒狒中获得。 为了这 目的，一种特定的单克隆抗体，仅识别 氧化抑制剂将用作主要染色试剂， 随后建立二抗免疫组化 方法（过氧化物酶-抗过氧化物酶、抗生物素蛋白-生物素）。 这个目标 对人类健康很重要，因为 α1- 的氧化 吸烟导致肺部蛋白酶抑制剂 是肺病的主要致病因素 肺气肿，但尚未得到证实。 2）我们将探讨的作用 新发现的肥大细胞弹性蛋白酶在发病机制中的作用 吸烟者的肺气肿可以通过三种方式：（a）通过监测释放 来自纯化人肺的酶（使用 ELISA 技术） 肥大细胞与烟草烟雾的水溶液一起孵育 烟雾成分； (b) 通过测量组胺的水平（组胺的标志物） 支气管肺泡灌洗液中的肥大细胞分泌反应 人类吸烟者（吸烟前和吸烟后）和老鼠的吸烟者 急性暴露于吸入香烟烟雾； (c) 通过尝试 在大鼠和狗中诱导实验性肺气肿 肥大细胞促分泌剂反复肺内滴注。 3）我们将尝试开发新的人类中性粒细胞抑制剂 弹性蛋白酶（一种被怀疑为致病因素的蛋白酶 肺气肿）基于我们最近的特征 肺炎球菌中的特异性中性粒细胞弹性蛋白酶抑制剂。 4）我们会 测试抗氧化、基因工程突变体（Met 358Val) α1-蛋白酶抑制剂，因其具有抗性 在已建立的急性急性白血病动物模型中体内失活 香烟烟雾暴露。 5）最后，我们将测量 血液中的中性粒细胞标记抗原 (L1) 和 有感染风险的脓毒症和创伤患者的气腔分泌物 成人呼吸窘迫综合征 (ARDS) 和患有以下疾病的患者 坦率的ARDS。 L1 蛋白的水平可能被证明具有 ARDS 对高危受试者的预测价值。