MOLECULAR BASIS FOR MICROBICIDAL ACTION IN LEUKOCYTES

白细胞杀菌作用的分子基础

• 批准号: 3126450
• 负责人: JAMES K HURST
• 金额: $ 14.81万
• 依托单位: OREGON GRADUATE INSTITUTE SCIENCE & TECH
• 依托单位国家: 美国
• 财政年份: 1979
• 资助国家: 美国
• 起止时间: 1979-04-01 至 1989-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3126450
• 关键词: bactericidal immunity; bacteriolysis; bioenergetics; cellular immunity; free radicals; hydroxyl group; ion transport; mass spectrometry; neutrophil; nucleoside diphosphate kinase; nucleotides; oxidation; oxidizing agents; peroxidases; phagocytosis; toxin

Studies are proposed to identify oxidative reactions occurring in phagocytosing leukocytes that are associated with microbicidal action. We have shown that the oxidant, hypochlorous acid (HOC1), inactivates E. coli in reactions that lead to destruction of metabolic energy reserves within the cell and to loss of energy-linked functions such as active transport. To explore the underlying molecular mechanisms, membrane localized energy-linked systems, including the ATP synthetase and selected ion and metabolite transport proteins, will be examined for evidence of oxidative damage. Studies to identify the lethal reactions of hydroxyl radical (OH) or OH-radical-generating systems willl be initiated. Comparison will be made between cell viabilities and loss of specific metabolic functions or biochemical components to identify loci of oxidative attack associated with cellular death. For this purpose differing methods of forming hydroxyl radical will be used that allow its generation in the extracellular medium, at the cell usrface, or within the microbial cell. Following protocols developed for study of inactivation by HOC1, the consequences of OH oxidation upon cellular transport systems, nucleotide concentrations, protonmotive force, and respiratory components (when appropriate) will be determined. For both HOC1 and OH oxidations, various organisms with widely differing metabolic requirements and cellular topographic organization will be studied to determine if particular inactivation mechanisms are "universal" in character. Organisms exhibiting clearly different mechanisms of oxidative inactivation by HOC1 and OH will be used to identify the toxin produced in neutrophil phagosomes. For these experiments the microbes will be phagocytosed, then immediately recovered to determine the cause of death. These studies should greatly clarify the nature of leukocytic mechanisms comprising host response to infection.

提出了研究以确定发生在 吞噬与微生物作用相关的白细胞。 我们 已经表明氧化剂次伐多酸（HOC1）使大肠杆菌失活 在导致代谢能量储量破坏的反应中 细胞和能量连锁函数的丧失，例如主动运输。 为了探索潜在的分子机制，膜本地化 能量连接系统，包括ATP合成酶和选定的离子和 代谢物转运蛋白将检查是否有氧化的证据 损害。 研究识别羟基自由基的致命反应（OH） 或将启动OH-RADICAL-RADICAL-GENERARY SYSTEMPL。 比较将是 在细胞的可行性和特定代谢功能的丧失之间或 鉴定与氧化攻击的基因座的生化成分 细胞死亡。 为此，形成羟基的不同方法 将使用激素，使其在细胞外介质中产生， 在细胞Usrface或微生物细胞内。 以下协议 开发用于通过HOC1灭活的研究，OH的后果 细胞运输系统上的氧化，核苷酸浓度， 质子运动力和呼吸部件（如果适当）将是 决定。 对于HOC1和OH氧化，各种生物都广泛 不同的代谢需求和蜂窝地形组织将 研究以确定特定的失活机制是否是 性格“通用”。 有机体明显不同 HOC1和OH的氧化灭活机制将用于 鉴定中性粒细胞中产生的毒素。 为此 实验将吞噬微生物，然后立即恢复 确定死亡原因。 这些研究应该大大阐明 白细胞机制的性质包括宿主对感染的反应。