METABOLIC ASPECTS OF BACTERIAL ECOLOGY IN HOST TISSUE

人中性粒细胞多形粒细胞（PMN）位于第一线 抵抗感染的细胞防御系统 它们吞噬细菌并杀死 它们具有氧依赖性或氧 独立的 中性粒细胞的颗粒是这些活动所固有的。 我们 我们的目标是继续确定蛋白质，占抗菌 粗颗粒提取物的活性。 我们的重点将放在 抗变形杆菌活性和纯化的阳离子抗微生物蛋白（CAP）， CAP 57和CAP 37对沙门氏菌和大肠杆菌有杀灭作用。杆菌 我们将：定义基因 和（组织）环境因素导致敏感性或 微生物的抗性;确定CAP相互作用的靶分子 与敏感细菌的外膜结合。 我们将完成研究 在细胞和亚细胞水平定位CAP。 我们将使用 了解细菌致病机理和宿主所需的技术 阻力 我们将用途：细胞培养、分化和梯度 离心;吞噬作用;抗菌作用测定;蛋白质 净化，包括常规和高性能液体 层析和亲和层析; ELISA;单克隆抗体; 通过P22转导构建等基因菌株;和 沙门氏菌的脂多糖和外膜蛋白的分析 它们对CAP的抗性不同。 我们的长期目标是确定CAP在氧非依赖性 抗微生物吞噬作用，并开发单克隆抗体快速 临床感染性疾病中中性粒细胞的筛查 我们提出的研究 它的重要性在于：a）确定至少存在 三种高效的阳离子抗菌蛋白（CAPs）， 在人PMN中的作用不依赖于氧; B）提供生化证据 对CAP作用敏感的细菌上的特定靶位点; c） 表明低氧张力对N. 淋病; d）提供高度暗示的证据，证明存在 迄今未被怀疑的抗微生物颗粒。

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.