title: Activation of phospholipase D by interleukin-8 in human neutrophils.
abstract: Interleukin 8 (IL-8), a member of the C-X-C branch of the chemokine superfamily, stimulated the breakdown of 1-O-[3H]alkyl-2-acyl-sn-glycero-3-phosphocholine ([3H]EAPC) and the formation of 1-O-[3H]alkyl-2-acyl-phosphatidic acid ([3H]-EAPA) in human polymorphonuclear leukocytes (PMN) in the presence of cytochalasin B. In addition, the mass of diradyl-PA was increased with similar kinetics. In the presence of ethanol, 1-O-[3H]alkyl-2-acyl-phosphatidylethanol ([3H]EAPEt) was formed at the expense of [3H]EAPA formation, indicating the activation of phospholipase D by the cytokine. The effect was time- and concentration-dependent, reaching a plateau at 30 seconds with the maximally activating concentration of 120 nmol/L IL-8. Preincubation of cells with 1 microgram/mL Bordetella pertussis toxin inhibited the breakdown of [3H]EAPC and [3H]EAPA formation, indicating a role for a pertussis toxin-sensitive guanosine triphosphate-binding protein. Formation of phosphatidic acid (PA) correlated with activation of nicotinamide adenine dinucleotide phosphate (NADPH) oxidase, the oxidative burst enzyme, with both events occurring in the same concentration range. Inhibition of PA formation, by the presence of ethanol, also inhibited the oxidative burst stimulation by IL-8. Pretreatment of PMN with 10 nmol/L platelet-activating factor potentiated both [3H]EAPA accumulation and activation of NADPD oxidase by IL-8. Collectively, these data show that IL-8 stimulates the metabolism of choline-containing phosphoglycerides in human PMN and support a role for PA in the signaling mechanisms used by IL-8 to stimulate PMN function.
authors.affiliation: Istituto di Ricerche Farmacologiche Mario Negri, Milan, Italy.
authors.forename: S
authors.initials: S
authors.lastname: Sozzani
authors.forename: D E
authors.initials: DE
authors.lastname: Agwu
authors.forename: M D
authors.initials: MD
authors.lastname: Ellenburg
authors.forename: M
authors.initials: M
authors.lastname: Locati
authors.forename: M
authors.initials: M
authors.lastname: Rieppi
authors.forename: A
authors.initials: A
authors.lastname: Rojas
authors.forename: A
authors.initials: A
authors.lastname: Mantovani
authors.forename: L C
authors.initials: LC
authors.lastname: McPhail
mesh_terms: D004305:Dose-Response Relationship, Drug; D004789:Enzyme Activation / Q000187:drug effects; D016209:Interleukin-8 / Q000037:antagonists & inhibitors / Q000494:pharmacology*; D009247:NADH, NADPH Oxidoreductases / Q000378:metabolism; D019255:NADPH Oxidases; D009504:Neutrophils / Q000187:drug effects*; D037342:Pertussis Toxin; D010739:Phospholipase D / Q000378:metabolism*; D010972:Platelet Activating Factor / Q000031:analogs & derivatives / Q000378:metabolism / Q000494:pharmacology; D016897:Respiratory Burst / Q000187:drug effects; D013481:Superoxides / Q000378:metabolism; D010566:Virulence Factors, Bordetella / Q000494:pharmacology
chemical_list: C036761:1-alkyl-2-acyl-sn-glycero-3-phosphocholine; D016209:Interleukin-8; D010972:Platelet Activating Factor; D010566:Virulence Factors, Bordetella; D013481:Superoxides; D009247:NADH, NADPH Oxidoreductases; D019255:NADPH Oxidases; D037342:Pertussis Toxin; D010739:Phospholipase D