ANTIMICROBIAL PEPTIDES OF HUMAN NEUTROPHILS

人类中性粒细胞的抗菌肽

• 批准号: 3134420
• 负责人: ROBERT IRVING LEHRER
• 金额: $ 17.12万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 1985
• 资助国家: 美国
• 起止时间: 1985-12-01 至 1988-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3134420
• 关键词: Bacteroides; Candida albicans; Clostridium; Cryptococcus neoformans; Escherichia coli; Pseudomonas aeruginosa; Salmonella typhimurium; Staphylococcus aureus; Vesiculovirus; aerobic bacteria; alveolar macrophages; anaerobic bacteria; antibacterial agents; antibiotics; antifungal agents; bactericidal immunity; enzyme linked immunosorbent assay; gel electrophoresis; gel filtration chromatography; gram negative bacteria; gram positive bacteria; herpes simplex virus 1; high performance liquid chromatography; human subject; immunochemistry; ion exchange chromatography; lysozyme; microorganism immunology; myeloperoxidase; neutrophil; peptides; phagocytosis; secondary infection

The long term objective of this research is to define in biochemical terms the microbicidal mechanisms of phagocytes in order to better understand the causes of opportunistic infection and to seek new means for its prevention and therapy. With their ability to ingest and kill potential pathogens, human neutrophils (PMN) are key components of host defenses against infection. We recently discovered that human PMN contain three antimicrobial peptides, named HNP-1, HNP-2 and HNP-3. The peptides, collectively referred to as "defensins", were sequenced. They exhibit homology to a family of six antimicrobial peptides abundant in rabbit granulocytes and/or lung macrophages, the "lysosomal cationic proteins" of Zeya and Spitznagel. The specific aims are as follows: a. to determine the antimicrobial spectrum of the human defensins by studying their ability to kill or inactivate aerobic and anaerobic bacteria, yeast-phase fungi and selected viruses. b. to identify and analyse factors that modulate the antimicrobial activity of defensins. c. to examine the interactions of these peptides with other potentially antimicrobial constituents of human neutrophils, including lysozyme, BPI, myeloperoxidase and selected reactive oxygen intermediates. d. to establish the intracellular location of defensins in PMN and assess both their intracellular translocation to phagolysosomes and their extracellular release. e. to ascertain the mechanisms whereby defensins act on microorganisms. f. to develop and apply qualitative and quantitative procedures to measure the defensin content of normal PMN and determine whether PMN defensin deficiency underlies the occurrence of certain, presently unexplained infections in man. The methodology for performing these studies includes cellular fractionation, chromatographic purification and various electrophoretic, immunocytochemical and immunoquantitative (ELISA) techniques. Specific details and examples are provided in the body of the proposal.

这项研究的长期目标是用生化术语定义 吞噬细胞的杀生机制，以便更好地了解 机会性感染的原因，并寻求新的手段以进行预防 和治疗。 人类摄入和杀死潜在病原体的能力 中性粒细胞（PMN）是宿主防御感染的关键组成部分。 我们最近发现，人PMN包含三个抗菌肽， 名为HNP-1，HNP-2和HNP-3。 肽，共同称为 测序“防御素”。 他们与六口家庭表现出同源 兔粒细胞中丰富的抗菌肽和/或肺 巨噬细胞，Zeya和Spitznagel的“溶酶体阳离子蛋白”。 具体目的如下：确定抗菌剂 人类防御素的频谱通过研究其杀戮能力或 灭活有氧和厌氧菌，酵母菌真菌并选择 病毒。 b。识别和分析调节的因素 防御素的抗菌活性。 c。检查的相互作用 这些具有人类其他潜在抗菌成分的肽 中性粒细胞，包括溶菌酶，BPI，髓过氧化物酶和选定的反应性 氧中间体。 d。建立细胞内位置 PMN中的防御素，并评估其细胞内易位 吞噬物体及其细胞外释放。 e。确定 防御素对微生物作用的机制。 f。发展和 应用定性和定量程序来测量防御素 正常PMN的含量并确定PMN防御能力缺陷是否缺乏 基础是某些目前无法解释的感染 男人。 进行这些研究的方法包括细胞 分馏，色谱纯化和各种电泳， 免疫细胞化学和免疫定量（ELISA）技术。 具体的 提案正文中提供了详细信息和示例。