P. GINGIVALIS MEMBRANE BLEBS-MODULATORS OF PMN FUNCTION

厌氧菌经常从临床感染中恢复 在整个身体。 拟杆菌属物种数量增加， 一种多微生物的植物群，通常与 慢性化脓性感染。 拟杆菌属物种诱导 混合感染的慢性化已得到证实，但尚不清楚 尽管有人认为它们可能影响宿主的防御 细胞，主要是多形核白细胞（PMN）。 进步 破坏性牙周病变含有数量增加的卟啉单胞菌 牙龈拟杆菌（以前的牙龈拟杆菌），我们假设， 微生物通过影响PMN而对疾病进展发挥作用 在牙周环境中发挥作用。 我们已经证明， 牙龈卟啉单胞菌的亲本菌株影响PMN功能，而其它菌株不影响。 PMN的调节活性位于外膜， 暴露在重封装菌株中。 然而，菌株释放出许多 膜泡进入细胞外环境和我们的初步研究 已经表明，以前的“PMN-非反应性”亲本菌株释放“PMN- 反应性”膜泡。 由于生理压力时水泡释放会增加，并且可能 与牙周炎的爆发相吻合，并成为牙周炎爆发的重要指标 本提案的具体目标是：（1） 表征对人体的生理和生化影响 外周血PMN牙龈卟啉单胞菌泡和外膜蛋白，（2） 鉴定在大泡和外膜中PMN调节活性的存在， 先前表征的“PMN非反应性”菌株的膜，和（3） 分离和表征负责任的外膜组分， P.牙龈炎 我们将描述PMN从初始膜 去极化通过动员细胞内第二信使， 呼吸爆发、脱粒和PMN运动的激活。 这些试验将根据趋化性和非趋化性进行。 趋化刺激和膜对反应的调节 将评价水泡和外膜蛋白。 PMN调节 将检查并比较大泡和外膜的活性 各种先前表征的“PMN-反应性”组分， “中性粒细胞无反应性”亲本菌株。 将分离PMN调节分子 使用Zwittergent洗涤剂提取细菌表面成分 然后进行色谱和电泳分离和纯化， 使用PMN测定作为生物活性的测试。

Anaerobic bacteria are frequently recovered from clinical infections throughout the body. Increased numbers of Bacteroides species, as part of a polymicrobial flora, are most often associated with the presence of chronic, purulent infections. The ability of Bacteroides species to induce chronicity to mixed infection has been demonstrated but not understood although it has been suggested that their effect may be on host defense cells, primarily the polymorphonuclear leukocyte (PMN). Progressive destructive periodontal lesions contain increased numbers of Porphyromonas gingivalis (formerly Bacteroides gingivalis) and we hypothesize that this microorganism exerts its effect on disease progression by affecting PMN function in the periodontal environment. We have demonstrated that some parent strains of P. gingivalis affect PMN function whereas others do not. The PMN-modulating activity located in the outer membrane and may not be exposed in heavily encapsulated strains. However, strains release numerous membrane blebs into the extracellular environment and our initial studies have shown that previously "PMN-unreactive" parent strains release "PMN- reactive" membrane blebs. Since bleb release is increased at times of physiologic stress and may coincide with, and be a important indicator of, bursts of periodontal destruction, the specific aims of this proposal are directed towards (1) characterizing the physiological and biochemical effects on human peripheral blood PMN P. gingivalis blebs and outer membrane proteins, (2) identifying the presence of PMN-modulating activity in the blebs and outer membranes of previously characterized "PMN-unreactive" strains, and (3) isolating and characterizing the responsible outer membrane components of P. gingivalis. We will characterize the PMN response from initial membrane depolarization through mobilization of intracellular second messengers, to activation of the respiratory burst, degranulation and PMN locomotion. These assays will be performed in response to chemotactic and non- chemotactic stimulation and the modulation of the responses by membrane blebs and outer membrane proteins will be evaluated. PMN-modulating activity will be examined and compared for blebs and outer membrane components of a variety of previously characterized "pmn-reactive" and "PMN-unreactive" parent strains. PMN-modulating molecules will be isolated using Zwittergent detergent extraction of the bacterial surface components followed by chromatographic and electrophoretic separation and purification using PMN assays as a test for biological activity.