MOLECULAR BASIS FOR MICROBICIDAL ACTION IN LEUKOCYTES

白细胞杀菌作用的分子基础

• 批准号: 2003194
• 负责人: JAMES K HURST
• 金额: $ 5.87万
• 依托单位: WASHINGTON STATE UNIVERSITY
• 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 2001-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2003194
• 关键词: Candida albicans; Escherichia coli; Raman spectrometry; Saccharomyces cerevisiae; bactericidal immunity; bioenergetics; carbonates; cell membrane; cellular immunity; chlorine; free radical oxygen; glucose metabolism; hydrogen peroxide; hydroxyl radical; leukocytes; membrane potentials; neutrophil; nitric oxide; oxidation reduction reaction; oxidizing agents; phagocytosis; stop flow technique; tissue /cell culture; toxin; vesicle /vacuole

DESCRIPTION (Adapted from applicant's abstract): Research is proposed to: 1) use specifically designed probes to identify the toxins generated by stimulated phagocytes and to monitor their intracellular reaction. 2) characterize their oxidation-reduction chemistry in sufficient detail that their potential role in cellular reactions can be properly evaluated, and 3) investigate the microbicidal mechanisms of putative phagocyte-generated toxins on representative test organisms. The oxidants include HOCl, H2O2, ONO2- and secondary oxidants derived from them. Experiments will use stopped-flow kinetics, esr spin trapping and Raman spectroscopy to identify the reaction products and to characterize their behavior toward biological materials. Recent studies have demonstrated that lethal reactions of very reactive oxidants cannot be accurately assessed by in vitro microbicidal assay systems. Fluorescein-conjugated polyacrylamide beads which are capable of discriminating between chlorine-based oxidants, ONO2- ion, and carbon-based oxidants will be used to probe events in the phagocytic vacuole. The bactericidal mechanisms of these oxidants will be examined by the protocols used for HOCl-inflicted cell damage. The studies will focus on metabolic functions associated with the bacterial plasm membrane. Cellular studies will use control neutrophils, a macrophage cell line, and MPO-deficient and CGD neutrophils.

描述（根据申请人的摘要改编）：提出研究至： 1）使用专门设计的探针来识别由 刺激吞噬细胞并监测其细胞内反应。 2） 以足够详细的详细表征其氧化化学的化学 可以正确评估它们在细胞反应中的潜在作用，3） 研究假定的吞噬细胞生成的微生物机制 代表性测试生物的毒素。 氧化剂包括HOCL，H2O2， 源自它们的Ono2和次要氧化剂。 实验将使用 停止流动动力学，ESR旋转诱捕和拉曼光谱学识别 反应产物并表征其对生物学的行为 材料。 最近的研究表明，非常 反应性氧化剂不能通过体外微生物准确评估 测定系统。 荧光素偶联的聚丙烯酰胺珠 能够区分基于氯的氧化剂，ono2-离子和 碳基氧化剂将用于探测吞噬细胞中的事件 液泡。 这些氧化剂的杀菌机制将通过 用于HOCL造成的细胞损伤的方案。 研究将集中 在与细菌浆膜相关的代谢功能上。 细胞研究将使用对照中性粒细胞，巨噬细胞系和 MPO缺陷和CGD中性粒细胞。