Role of Pseudomonas aeruginosa lipase in inflammatory mediator release from human inflammatory effector cells (platelets, granulocytes, and monocytes.
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.
König
B
B
Medizinische Mikrobiologie und Immunologie, AG Infektabwehrmechanismen, Ruhr-Universität Bochum, Germany.
Jaeger
K E
KE
Sage
A E
AE
Vasil
M L
ML
König
W
W
D019377:12-Hydroxy-5,8,10,14-eicosatetraenoic Acid; D001491:Basophils / Q000187:drug effects; D001792:Blood Platelets / Q000187:drug effects*; D002634:Chemotaxis, Leukocyte; D004798:Enzymes / Q000378:metabolism; D006632:Histamine / Q000032:analysis; D006801:Humans; D006893:Hydroxyeicosatetraenoic Acids / Q000032:analysis; D007249:Inflammation / Q000378:metabolism*; D016209:Interleukin-8 / Q000032:analysis; D007962:Leukocytes / Q000187:drug effects*; D007975:Leukotriene B4 / Q000032:analysis; D008049:Lipase / Q000494:pharmacology*; D008163:Luminescent Measurements; D009000:Monocytes / Q000187:drug effects; D009504:Neutrophils / Q000187:drug effects; D011550:Pseudomonas aeruginosa / Q000201:enzymology*; D010738:Type C Phospholipases / Q000494:pharmacology
D004798:Enzymes; D006893:Hydroxyeicosatetraenoic Acids; D016209:Interleukin-8; D007975:Leukotriene B4; D019377:12-Hydroxy-5,8,10,14-eicosatetraenoic Acid; D006632:Histamine; D008049:Lipase; D010738:Type C Phospholipases