Role of Pseudomonas aeruginosa lipase in inflammatory mediator release from human inflammatory effector cells (platelets, granulocytes, and monocytes.
Role of Pseudomonas aeruginosa lipase in inflammatory mediator release from human inflammatory effector cells (platelets, granulocytes, and monocytes.
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铜绿假单胞菌脂肪酶在人类炎症效应细胞（血小板、粒细胞和单核细胞）释放炎症介质中的作用。
DOI：
10.1128/iai.64.8.3252-3258.1996
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发表时间：
1996
影响因子：
3.1
通讯作者：
König,W
中科院分区：
文献类型：
--
作者：
König,B;Jaeger,KE;Sage,AE;Vasil,ML;König,W
先前，我们已经证明铜绿假单胞菌脂肪酶和磷脂酶C （PLC）是两种细胞外脂解酶，在人血小板生成12-羟基二碳四烯酸（HETE）过程中相互作用。在这方面。将纯化的铜绿假单胞菌脂肪酶添加到含有plc的铜绿假单胞菌粗培养上清液中，可以提高人血小板中化学活性12-HETE的生成。因此，我们分析了纯化的铜绿假单胞菌脂肪酶和纯化的溶血性铜绿假单胞菌PLC在人血小板、嗜中性粒细胞和嗜碱性粒细胞以及单核细胞中炎症介质释放方面的相互作用。纯化的P. aeruginosa PLC，但本身没有纯化脂肪酶，诱导人血小板产生12-HETE，白三烯B4 （LTB4）和氧代谢产物的产生，人中性粒细胞释放酶，嗜碱性粒细胞释放组胺，但以剂量依赖性方式减少人单核细胞释放白细胞介素-8 （IL-8）。添加纯化脂肪酶增强了plc诱导的12-HETE和LTB4的生成，不影响酶、组胺或IL-8的释放，但降低了plc诱导的化学发光反应。用产气荚膜梭菌的溶血性PLC代替铜绿假单胞菌的PLC也得到了类似的结果。为了进一步比较，我们使用定义明确的钙离子载体A23187和磷酸氢酶-12-肉豆酸-13-乙酸酯（PMA）作为刺激。脂肪酶增强了钙离子载体诱导的LTB4生成和β -葡糖醛酸酶释放，但减少了钙离子载体诱导和pma诱导的化学发光。同时，我们分析了脂肪酶在含PLC和脂肪酶的铜绿假单胞菌粗培养上清中的作用。P. aeruginosa培养上清液中的脂肪酶活性被脂肪酶特异性抑制剂十六烷基磺酰氟（hexadecylsulfonyl fluoride）抑制，PLC活性不受影响。“脂肪酶灭活培养上清”诱导12-HETE和LTB4生成的能力降低了50% ~ 100%。我们的研究结果表明，寄生在被感染宿主体内的铜绿假单胞菌同时分泌脂肪酶和PLC可能导致严重的病理影响，而这种影响不能用单个毒力因子对炎症效应细胞的单一作用来解释。
Previously, we have shown that Pseudomonas aeruginosa lipase and phospholipase C (PLC), two extracellular lipolytic enzymes, interact with each other during 12-hydroxyeicosatetraenoic acid (HETE) generation from human platelets. In this regard. the addition of purified P. aeruginosa lipase to PLC-containing crude P. aeruginosa culture supernatants enhances the generation of the chemotactically active 12-HETE from human platelets. Therefore, we analyzed the interaction of purified P. aeruginosa lipase and purified hemolytic P. aeruginosa PLC with regard to inflammatory mediator release from human platelets, neutrophilic and basophilic granulocytes, and monocytes. Purified P. aeruginosa PLC, but not purified lipase by itself, induced 12-HETE generation from human platelets, the generation of leukotriene B4 (LTB4) and oxygen metabolites, enzyme release from human neutrophils, and histamine release from basophils but diminished interleukin-8 (IL-8) release from human monocytes in a dose-dependent manner. The addition of purified lipase enhanced PLC-induced 12-HETE and LTB4 generation, did not influence enzyme, histamine, or IL-8 release, but diminished the PLC-induced chemiluminescent response. Similar results were obtained when the hemolytic PLC from Clostridium perfringens was used instead of P. aeruginosa PLC. For further comparison, we used the well-defined calcium ionophore A23187 and phorbol-12-myristate-13-acetate (PMA) as stimuli. Lipase enhanced calcium ionophore-induced LTB4 generation and beta-glucuronidase release but reduced calcium ionophore-induced and PMA-induced chemiluminescence. In parallel, we analyzed the role of lipase in a crude P. aeruginosa culture supernatant containing PLC and lipase. Lipase activity in the P. aeruginosa culture supernatant was inhibited by treatment with the lipase-specific inhibitor hexadecylsulfonyl fluoride, leaving the activity of PLC unaffected. The capacity of "lipase-inactivated culture supernatant" to induce 12-HETE and LTB4 generation was diminished by 50 to 100%. Our results suggest that the simultaneous secretion of lipase and PLC by P. aeruginosa residing in an infected host may result in severe pathological effects which cannot be explained by the sole action of the individual virulence factor on inflammatory effector cells.