title: Esterified eicosanoids are acutely generated by 5-lipoxygenase in primary human neutrophils and in human and murine infection.
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/10(6) 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 Ca(2+), phospholipase C, cytosolic and secretory phospholipase A(2), 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.
authors.affiliation: Department of Infection, Immunity and Biochemistry, School of Medicine, Cardiff University, Cardiff, United Kingdom.
authors.forename: Stephen R
authors.initials: SR
authors.lastname: Clark
authors.forename: Christopher J
authors.initials: CJ
authors.lastname: Guy
authors.forename: Martin J
authors.initials: MJ
authors.lastname: Scurr
authors.forename: Philip R
authors.initials: PR
authors.lastname: Taylor
authors.forename: Ann P
authors.initials: AP
authors.lastname: Kift-Morgan
authors.forename: Victoria J
authors.initials: VJ
authors.lastname: Hammond
authors.forename: Christopher P
authors.initials: CP
authors.lastname: Thomas
authors.forename: Barbara
authors.initials: B
authors.lastname: Coles
authors.forename: Gareth W
authors.initials: GW
authors.lastname: Roberts
authors.forename: Matthias
authors.initials: M
authors.lastname: Eberl
authors.forename: Simon A
authors.initials: SA
authors.lastname: Jones
authors.forename: Nicholas
authors.initials: N
authors.lastname: Topley
authors.forename: Sailesh
authors.initials: S
authors.lastname: Kotecha
authors.forename: Valerie B
authors.initials: VB
authors.lastname: O'Donnell
mesh_terms: D000368:Aged; D000369:Aged, 80 and over; D000818:Animals; D001094:Arachidonate 5-Lipoxygenase / Q000378:metabolism*; D001424:Bacterial Infections / Q000378:metabolism*; D015777:Eicosanoids / Q000096:biosynthesis* / Q000737:chemistry; D005260:Female; D016908:Gram-Positive Bacterial Infections / Q000378:metabolism; D006801:Humans; D006893:Hydroxyeicosatetraenoic Acids / Q000096:biosynthesis / Q000737:chemistry; D066298:In Vitro Techniques; D016209:Interleukin-8 / Q000096:biosynthesis; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008875:Middle Aged; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000494:pharmacology; D009504:Neutrophils / Q000187:drug effects / Q000378:metabolism*; D010538:Peritonitis / Q000378:metabolism; D010743:Phospholipids / Q000096:biosynthesis / Q000737:chemistry; D010955:Plasmalogens / Q000096:biosynthesis / Q000737:chemistry; D015398:Signal Transduction; D013203:Staphylococcal Infections / Q000378:metabolism; D013212:Staphylococcus epidermidis; D013481:Superoxides / Q000378:metabolism; D053719:Tandem Mass Spectrometry; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
chemical_list: C508606:CXCL8 protein, human; D015777:Eicosanoids; D006893:Hydroxyeicosatetraenoic Acids; D016209:Interleukin-8; D010743:Phospholipids; D010955:Plasmalogens; D013481:Superoxides; C022022:5-hydroxy-6,8,11,14-eicosatetraenoic acid; D009240:N-Formylmethionine Leucyl-Phenylalanine; D001094:Arachidonate 5-Lipoxygenase; D013755:Tetradecanoylphorbol Acetate