1. Esterified eicosanoids are acutely generated by 5-lipoxygenase in primary human neutrophils and in human and murine infection
- Author
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Christopher P. Thomas, Victoria Jayne Hammond, Sailesh Kotecha, Martin J. Scurr, Christopher J. Guy, Simon Arnett Jones, Stephen Clark, Barbara Coles, Gareth Roberts, Nicholas Topley, Matthias Eberl, Ann Kift-Morgan, Philip R. Taylor, and Valerie B. O'Donnell
- Subjects
Male ,Neutrophils ,Biochemistry ,Mice ,Phagocytes, Granulocytes, and Myelopoiesis ,chemistry.chemical_compound ,0302 clinical medicine ,Superoxides ,Tandem Mass Spectrometry ,Staphylococcus epidermidis ,Hydroxyeicosatetraenoic Acids ,Cytochalasin ,Phospholipids ,Aged, 80 and over ,0303 health sciences ,biology ,Superoxide ,Bacterial Infections ,Hematology ,Middle Aged ,Staphylococcal Infections ,N-Formylmethionine Leucyl-Phenylalanine ,Arachidonate 5-lipoxygenase ,cardiovascular system ,Tetradecanoylphorbol Acetate ,Female ,lipids (amino acids, peptides, and proteins) ,Signal Transduction ,Bacterial Peritonitis ,Plasmalogens ,Immunology ,In Vitro Techniques ,Peritonitis ,Microbiology ,03 medical and health sciences ,Phosphatidylcholine ,Animals ,Humans ,Gram-Positive Bacterial Infections ,Aged ,030304 developmental biology ,Phosphatidylethanolamine ,Arachidonate 5-Lipoxygenase ,Interleukin-8 ,Cell Biology ,Neutrophil extracellular traps ,biology.organism_classification ,Mice, Inbred C57BL ,chemistry ,biology.protein ,Eicosanoids ,030217 neurology & neurosurgery - Abstract
5-Lipoxygenase (5-LOX) plays key roles in infection and allergic responses. Herein, four 5-LOX–derived lipids comprising 5-hydroxyeicosatetraenoic acid (HETE) attached to phospholipids (PLs), either phosphatidylethanolamine (PE) or phosphatidylcholine (18:0p/5-HETE-PE, 18:1p/5-HETE-PE, 16:0p/5-HETE-PE, and 16:0a/5-HETE-PC), were identified in primary human neutrophils. They formed within 2 minutes in response to serum-opsonized Staphylococcus epidermidis or f-methionine-leucine-phenylalanine, with priming by lipopolysaccharide, granulocyte macrophage colony-stimulating factor, or cytochalasin D. Levels generated were similar to free 5-HETE (0.37 ± 0.14 ng vs 0.55 ± 0.18 ng/106 cells, esterified vs free 5-HETE, respectively). They remained cell associated, localizing to nuclear and extranuclear membrane, and were formed by fast esterification of newly synthesized free 5-HETE. Generation also required Ca2+, phospholipase C, cytosolic and secretory phospholipase A2, 5-LOX activating protein, and mitogen-activated protein kinase/extracellular signal-regulated kinase kinase 1. 5-HETE-PLs were detected in murine S epidermidis peritonitis, paralleling neutrophil influx, and in effluent from Gram-positive human bacterial peritonitis. Formation of neutrophil extracellular traps was significantly enhanced by 5-LOX inhibition but attenuated by HETE-PE, whereas 5-HETE-PE enhanced superoxide and interleukin-8 generation. Thus, new molecular species of oxidized PL formed by human neutrophils during bacterial infection are identified and characterized.
- Published
- 2011
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