HUMAN PHAGOCYTES, OXYGEN METABOLITES AND INFLAMMATION

人类吞噬细胞、氧代谢物和炎症

• 批准号: 6510319
• 负责人: STEPHEN J WEISS
• 金额: $ 25.81万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 1984
• 资助国家: 美国
• 起止时间: 1984-07-01 至 2004-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6510319
• 关键词: adenosinetriphosphatase; carbohydrate receptor; cathepsin B; cellular pathology; chemoattractants; confocal scanning microscopy; cysteine endopeptidases; cystine; enzyme activity; extracellular matrix; high performance liquid chromatography; immunoelectron microscopy; inflammation; laboratory rabbit; lysosomes; mannose 6 phosphate; monoclonal antibody; oxidation; phagocytes; protease inhibitor; protein degradation; receptor binding; tissue /cell culture; western blottings

In chronic inflammatory disease states ranging from rheumatoid arthritis to ulcerative colitis, blood monocytes infiltrate affected tissues and differentiate into tissue-destructive populations of macrophages. Unlike neutrophils or eosinophils which predominantly rely on their ability to generate halogenated oxidants and release serine proteinases to mediate tissue damage, the destructive systems mobilized by human macrophages remain undefined. In order to identify and characterize the mechanisms by which macrophages mediate tissue-destructive effects in chronic inflammatory disease states, human monocytes will be cultured in vitro and induced to mature into a macrophage population capable of expressing an extracellular matrix-degrading phenotype hundreds of times greater than that of any other leukocyte population described previously. Based on preliminary data, the macrophage's unique destructive activity is mediated by the exocytosis/secretion of active cysteine proteinases, a class of acid proteases normally categorized as intracellular, lysosomal catabolic enzymes. To date, little is known with regard to the function of these enzymes in human macrophages since many members of this proteinase family have only recently been identified and few molecular or biochemical tools have been developed for their analysis in intact cell systems. Furthermore, the mechanisms by which lysosomal enzymes could be routed to an extracellular compartment in which conditions permissive for cysteine proteinase activity could be generated and maintained remain unknown. Thus, in an attempt to identify a novel role for cysteine proteinases in macrophage effector functions, the following five aims will be addressed. First, to characterize the intracellular and extracellular expression of the cysteine proteinases, cathepsins B, L and S, as well as the cysteine proteinase inhibitor, cystatin C, in monocyte-derived macrophages. Second, to determine the role of the mannose-6-phosphate receptor recognition systems in directing cysteine proteinase traffic from the lysosomal to the extracellular compartment. Third, to characterize the role of the macrophage vacuolar-type H+-ATPase and L-cystine transport systems in generating an acidic and reducing extracellular environment permissive for cysteine proteinase activity. Fourth, to assess the role of the cysteine proteinase system in the macrophage-mediated degradation of the extracellular matrix via the selective "knockout" of individual cathepsins, the vacuolar-type H+-- ATPase or the L-cystine transport system. Fifth, and finally, to determine the response and function of the macrophage cysteine proteinase system during chemotactic factor-induced tissue invasion. The characterization of the cysteine proteinase system mobilized by human macrophages at inflamed sites should not only provide new insights into the pathogenesis and treatment of chronic inflammatory disease states, but also into the regulation of the tissue-invasive phenotype in vivo.

处于类风湿性关节炎等慢性炎症疾病状态 对于溃疡性结肠炎，血单核细胞浸润受影响的组织并 分化为具有组织破坏性的巨噬细胞群。 不像 中性粒细胞或嗜酸性粒细胞主要依赖于它们的能力 产生卤化氧化剂并释放丝氨酸蛋白酶来介导 组织损伤，人类巨噬细胞动员的破坏系统 保持未定义。 为了识别和表征机制 巨噬细胞通过其介导慢性组织破坏作用 炎症性疾病状态下，将在体外培养人单核细胞 并诱导成熟为能够表达的巨噬细胞群 细胞外基质降解表型大数百倍 比之前描述的任何其他白细胞群的数量都要多。 基于 根据初步数据，巨噬细胞独特的破坏活性是 由活性半胱氨酸蛋白酶的胞吐作用/分泌介导， 通常分为细胞内、溶酶体的一类酸性蛋白酶 分解代谢酶。 迄今为止，人们对该功能知之甚少 这些酶在人类巨噬细胞中的存在，因为该酶的许多成员 蛋白酶家族最近才被鉴定，并且很少有分子或 已开发出用于在完整细胞中进行分析的生化工具 系统。 此外，溶酶体酶的机制 路由到细胞外室，其中条件允许 可以产生并维持半胱氨酸蛋白酶活性 未知。 因此，在尝试确定半胱氨酸的新作用时 蛋白酶在巨噬细胞效应功能中的作用，有以下五个目的 将得到解决。 首先，表征细胞内和 半胱氨酸蛋白酶、组织蛋白酶 B、L 和 S 以及半胱氨酸蛋白酶抑制剂半胱氨酸蛋白酶抑制剂 C 单核细胞来源的巨噬细胞。 二、确定角色 6-磷酸甘露糖受体识别系统引导半胱氨酸 蛋白酶从溶酶体到细胞外区室的运输。 三、表征巨噬细胞液泡型H+-ATP酶的作用 和 L-胱氨酸转运系统产生酸性和还原性 细胞外环境允许半胱氨酸蛋白酶活性。 第四，评估半胱氨酸蛋白酶系统在 巨噬细胞介导的细胞外基质降解 选择性“敲除”单个组织蛋白酶，即液泡型 H+-- ATP 酶或 L-胱氨酸转运系统。 第五，也是最后， 确定巨噬细胞半胱氨酸蛋白酶的反应和功能 趋化因子诱导的组织侵袭过程中的系统。 这 人类动员的半胱氨酸蛋白酶系统的表征 发炎部位的巨噬细胞不仅应该提供新的见解 慢性炎症性疾病的发病机制和治疗， 还涉及体内组织侵袭表型的调节。

项目成果

Oxygen, ischemia and inflammation.

缺氧、缺血和炎症。

• 发表时间: 1986
• 作者: Weiss; S J
• 通讯作者: S J

Pericellular mobilization of the tissue-destructive cysteine proteinases, cathepsins B, L, and S, by human monocyte-derived macrophages.

人单核细胞来源的巨噬细胞对组织破坏性半胱氨酸蛋白酶、组织蛋白酶 B、L 和 S 的细胞周动员。

• DOI: 10.1073/pnas.92.9.3849
• 发表时间: 1995-04-25
• 期刊: Proceedings of the National Academy of Sciences of the United States of America
• 影响因子: 11.1
• 作者: V. Reddy;Qing Yu Zhang;S. Weiss
• 通讯作者: S. Weiss

Singlet oxygen production by human eosinophils.

人类嗜酸性粒细胞产生单线态氧。

• 发表时间: 1988-07-15
• 期刊: The Journal of biological chemistry
• 作者: J. R. Kanofsky;H. Hoogl;R. Wever;S. Weiss
• 通讯作者: S. Weiss

Regulation of elastinolytic cysteine proteinase activity in normal and cathepsin K-deficient human macrophages.

正常和组织蛋白酶 K 缺陷的人巨噬细胞中弹性蛋白溶解半胱氨酸蛋白酶活性的调节。

• 发表时间: 2000-09-18
• 作者: Punturieri, A;Filippov, S;Allen, E;Caras, I;Murray, R;Reddy, V;Weiss, S J
• 通讯作者: Weiss, S J

Matrilysin-dependent elastolysis by human macrophages.

人类巨噬细胞的基质溶素依赖性弹性溶解。

• 发表时间: 2003-09-15
• 作者: Filippov, Sergey;Caras, Ingrid;Murray, Richard;Matrisian, Lynn M;Chapman Jr, Harold A;Shapiro, Steven;Weiss, Stephen J
• 通讯作者: Weiss, Stephen J