METABOLIC ASPECTS OF BACTERIAL ECOLOGY IN HOST TISSUE

人中性粒细胞(PMN)处于第一线 对感染的细胞防御能力。它们吞噬细菌并杀死 它们具有依赖氧气或氧气的抗菌活性 独立自主。PMN颗粒是这些活动所固有的。我们的 目标是继续确定与抗菌剂有关的蛋白质 粗颗粒提取物的活性。我们的主要重点将是 抗变形杆菌活性和纯化的阳离子抗微生物蛋白(CAP)， 能杀死沙门氏菌和大肠杆菌的CAP57和CAP37。我们将：定义基因 和(组织)环境因素导致敏感性或 微生物的抗药性；确定CAP相互作用的靶分子 在敏感细菌的外膜上。我们会完成学业的 在细胞和亚细胞级别本地化CAP。我们将使用 了解细菌致病机理和宿主所需的技术 抵抗。我们将使用：细胞培养和分化和梯度 离心法；吞噬作用；抗菌试验；蛋白质 净化，包括常规液体和高性能液体 层析和亲和层析；酶联免疫吸附试验；单抗； P22转导等基因菌株的构建及生化 沙门氏菌内毒素和外膜蛋白的分析 小鼠对CAP的抗性不同。 我们的长期目标是确定CAP在氧非依赖性中的作用 抗微生物吞噬作用及快速单抗的研制 中性粒细胞在临床感染性疾病中的筛查我们提出的研究 具有重大意义，因为它将：a)至少确定存在 三种高效阳离子抗菌蛋白(CAPS) 在人体PMN中的作用不依赖于氧；b)提供生化证据 针对CAP作用的敏感细菌上的特定靶点；c) 说明低氧压对N。 淋病；d)提供极具启发性的证据，证明存在一种 迄今未被怀疑的抗菌颗粒。

Human neutrophil polymorphonuclear granulocytes (PMN) are in the first line of cellular defense against infection. They phagocytize bacteria and kill them with antimicrobial activities that are oxygen dependent or oxygen independent. The granules of PMN are intrinsic to these activities. Our goal is to continue to define the proteins that account for antimicrobial activities of crude granule extracts. Our main emphasis will be on anti-Proteus activity and purified cationic antimicrobial proteins (CAP), CAP57 and CAP37 that kill Salmonella and E. coli. We will: define genetic and (tissue) environmental factors contributing to sensitivity or resistance of microorganisms; determine the target molecules CAP interact with in the outer membrane of sensitive bacteria. We will complete studies to localize CAPs at the cellular and subcellular levels. We will use techniques required to understand bacterial pathogenesis and host resistance. We will use: cell culture and differential and gradient centrifugation; phagocytosis; assays for antimicrobial action; protein purification, including conventional and high performance liquid chromatography and affinity chromatography; ELISA; monoclonal antibodies; construction of isogenic strains by P22 transduction; and biochemical analysis of lipopolysaccharides, and outer membrane proteins of Salmonella typhimurium which differ in their resistance to CAP. Our long term objective is to define the role of CAP in oxygen independent antimicrobial phagocytosis and to develop monoclonal antibodies for rapid screening of PMN in clinical infectious disease. The research we propose is significant in that it will: a) establish the existence of at least three highly effective Cationic Antimicrobial Proteins (CAPs) whose mode of action in human PMN is oxygen independent; b) provide biochemical evidence for specific target sites on susceptible bacteria to the action of CAP; c) implicate a contribution of low O2 tension to the resistance to CAPs of N. gonorrhoeae; d) provide highly suggestive evidence for the existence of a heretofore unsuspected antimicrobial granule.