MURINE MODEL OF X LINKED CHRONIC GRANULOMATOUS DISEASE

X连锁慢性肉芽肿病的小鼠模型

• 批准号: 2230001
• 负责人: Mary C Dinauer
• 金额: $ 20.64万
• 依托单位: INDIANA UNIV-PURDUE UNIV AT INDIANAPOLIS
• 依托单位国家: 美国
• 财政年份: 1994
• 资助国家: 美国
• 起止时间: 1994-07-01 至 1999-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2230001
• 关键词: bacterial pneumonia; cellular pathology; chronic granulomatous disease; dexamethasone; endopeptidases; free radical oxygen; genetic models; genetic strain; histology; immunocytochemistry; inflammation; interferon gamma; interleukin 1; laboratory mouse; leukocyte oxidative burst; lung injury; microscopy; oxidizing agents; peritonitis; phagocytes; phenotype; respiratory infections; sex linked trait; tumor necrosis factor alpha

Chronic granulomatous disease (CGD) is a recessive genetic disorder in which phagocytes cannot generate superoxide and other reactive oxidants due to mutations in different subunits of the respiratory burst oxidase (NADPH-oxidase) complex. The clinical syndrome is characterized by recurrent, severe bacterial and fungal infections and chronic inflammation with granuloma formation. Mutations in the X-Iinked gene for gp91(phox), the larger subunit of the oxidase cytochrome b., account for the majority of cases of COD (X-CGD). A murine model of X-CGD has now been developed, based on the use of gene targeting to disrupt the X-linked gp91(phox) gene in murine embryonic stem cells. The phenotype of the X-CGD mouse, as manifested in the inbred C57BL/6 strain, will be studied in this proposal. The frequency of spontaneous infectious and granulomatous complications in a specific pathogen free environment will be monitored, and the relative susceptibility of X-CGD mice to catalase-positive and catalase-negative microbes will be determined in experimental pneumonia and peritonitis. Other experiments will address the hypotheses that phagoycte-generated oxygen radicals are both important mediators of inflammatory tissue injury and at the same time essential for normal resolution of the inflammatory process. Acute and chronic injury will be induced in the skin or lung using either bacteria or non-infectious agents. The natural history of the inflammatory lesion will be compared between X-CGD and wild type mice. In some experiments, mice with a combined genetic deficiency of both the respiratory burst and neutrophil neutral proteases (bg/bg) will also be studied. Evaluation of the host response will include assessment of tissue injury by histologic analysis and by quantitation of edema formation, and characterization of the inflammatory cell infiltrate using histologic, morphometric, and immunohistochemical techniques. Experimental conditions which induce chronic foci of inflammation in X-CGD mice will be determined, and the efficacy of dexamethasone and interferon-gamma in ameliorating this process will be examined. This studies should provide new insight into the pathogenesis and treatment of CGD, and serve as a foundation for future investigations of these mice, including the impact of gene replacement therapy and the interaction of the X-CGD allele with other genetic defects influencing the inflammatory response. More broadly, the experiments described in this proposal should contribute to knowledge of the role of phagocyte oxidant production in acute and chronic inflammation in other clinical settings, and may suggest new approaches to treatment of these disorders.

慢性肉芽肿性疾病（CGD）是一种隐性遗传疾病 哪些吞噬细胞不能产生超氧化物和其他反应性氧化剂 由于呼吸道爆发氧化酶不同亚基的突变 （NADPH-氧化酶）复合物。临床综合征的特征是 经常性，严重的细菌和真菌感染以及慢性炎症 与肉芽肿形成。 GP91（PHOX）的X二联基因中的突变， 氧化酶细胞色素b。的较大亚基，占多数 COD病例（X-CGD）。现在已经开发了X-CGD的鼠模型 基于使用基因靶向破坏X连锁GP91（PHOX）基因 在鼠类胚胎干细胞中。 X-CGD鼠标的表型，AS 在此提案中将研究在近交C57BL/6菌株中的表现。 自发感染和肉芽肿并发症的频率 将监控特定的无病原体环境，并相对 X-CGD小鼠对过氧化氢酶阳性和过氧化氢酶阴性的敏感性 微生物将在实验性肺炎和腹膜炎中确定。 其他实验将解决吞噬生成的假设 氧自由基都是炎症组织损伤的重要介质 同时对于正常分辨炎症至关重要 过程。皮肤或肺部将诱发急性和慢性损伤 使用细菌或非感染药物。自然历史 将比较X-CGD和野生型小鼠之间的炎症病变。在 一些实验，具有两者的遗传缺陷的小鼠 呼吸爆发和中性粒细胞中性蛋白酶（Bg/bg）也将是 研究。宿主反应的评估将包括对组织的评估 通过组织学分析和通过降水的定量损伤， 使用组织学的表征炎性细胞浸润 形态计量学和免疫组织化学技术。实验条件 哪些诱导X-CGD小鼠炎症的慢性焦点将是 确定，地塞米松和干扰素伽马在 改善此过程将进行检查。这项研究应提供 对CGD的发病机理和治疗的新见解，并充当 未来对这些老鼠进行调查的基础，包括影响 基因替代疗法以及X-CGD等位基因与 其他遗传缺陷会影响炎症反应。更广泛地 本提案中描述的实验应有助于知识 吞噬细胞氧化剂在急性和慢性中的作用 在其他临床环境中的炎症，并可能提出新的方法 这些疾病的治疗。