ROLE OF PROTEASE IN INFECTION & INFLAMMATION OF COPD

描述(改编自申请人的摘要) 该项目将分析中性粒细胞/单核细胞丝氨酸蛋白酶的作用， 弹性蛋白酶、组织蛋白酶-G和蛋白水解酶-3，以及它们的特异性抑制物--α- 1-抗胰蛋白酶，特别是MNEI(单核/中性粒细胞弹性酶抑制物)，在 慢性阻塞性肺疾病的感染和炎症。需要评估的假设是 炎性肺液中过多的蛋白水解酶导致疾病 进展，不仅通过引起炎症和损伤，而且还 重要的是，通过靶向和摧毁先天的抗微生物肺部 防御分子。主要基于调查人员最近的发现， 研究人员假设，一种重要的先天防御分子 表面活性蛋白-A(SP-A)受这些酶的保护，并受到MNEI的保护。 调色素和激活剂，已知可增强摄取和清除 常驻肺巨噬细胞的微生物。一项针对小鼠的研究被提出。 中性粒细胞/单核细胞蛋白酶活性过高的模型，研究人员 将通过靶向mnei基因产生。Mnei-A def缺陷小鼠将是 与野生型小鼠相比，它们清除肺部感染的能力 肺炎链球菌和非分型流感嗜血杆菌，显著 慢性支气管炎的感染性病原体。炎症参数(中性粒细胞 内流、游离蛋白酶、SP-A水平、细胞因子、趋化因子)也将 比较一下。使用互补的人体体外系统，研究人员将 慢性阻塞性肺疾病患者炎性肺液(BAL上清液)与肺的比较 正常健康的非吸烟者和吸烟者的体液，以增强他们的能力 正常巨噬细胞对肺炎链球菌和流感嗜血杆菌的摄取。这个 研究人员还将检查外源蛋白酶对细菌的影响 摄取增强活性(包括SP-A)。阐明弹性蛋白酶的作用， 组织蛋白酶-G和蛋白酶-3在慢性阻塞性肺疾病的疾病进展中起重要作用， 在其他原因中，因为治疗方式现在是可行的，并且可以 如果潜在的病理过程是 明白了。

DESCRIPTION (adapted from the applicants' abstract) The project will analyze the role of the neutrophil/monocyte serine proteases, elastase, cathepsin-G and proteinase-3, and their specific inhibitors, alpha- 1-antitrypsin and especially MNEI (monocyte/neutrophil elastase inhibitor), in infection and inflammation of COPD. The hypothesis to be evaluated is that excess of the proteases in inflammatory lung fluids contributes to disease progression, not only by inducing inflammation and injury, but also importantly by targeting and destroying innate anti-microbial pulmonary defense molecules. Based largely on the investigators recent findings, the investigators hypothesize that an important innate defense molecule targeted by the proteases, and protected by MNEI, is surfactant protein-A (SP-A), an opsonin and activating agent, known to enhance uptake and clearance of microbes by resident pulmonary macrophages. Studies are proposed in a mouse model of excess neutrophil/monocyte protease activity, which the investigators will generate by targeting the mnei gene. Mnei-A def deficient mice will be compared to wildtype mice for their ability to clear pulmonary infections with Streptococcus pneumoniae and nontypeable Haemophilus influenzae, significant infectious agents in chronic bronchitits. Inflammatory parameters (neutrophil influx, free protease, SP-A levels, cytokines, chemokines) will also be compared. Using a complementary human in vitro system, the investigators will compare inflammatory lung fluids (BAL supernatants) of COPD patients with lung fluids of normal healthy non-smokers and smokers, for their ability to enhance uptake of S. pneumoniae and H. influenzae by normal macrophages. The investigators will examine also the effect of exogenous protease on bacterial uptake enhancing activity (including SP-A). Clarifying the role of elastase, cathepsin-G and proteinase-3 in disease progression of COPD is important, among other reasons, because therapeutic modalities are now feasible and could be delivered to the patients if the underlying pathological processes were understood.