title: Endotoxins stimulate neutrophil adhesion followed by synthesis and release of platelet-activating factor in microparticles.
abstract: Lipopolysaccharides and triacyl-cysteine-modified proteins of Gram-negative and positive organisms are potent endotoxins. Animal models show that the receptor for platelet-activating factor (PAF) is responsible for many of the deleterious effects of endotoxin, where regulated, localized PAF production localizes the inflammatory response. In contrast, biologically active analogs of PAF (PAF-like lipids) are generated by oxidative attack on phospholipids by chemical reactions that are unregulated and unlocalized. The identity and distribution of the PAF receptor ligand in endotoxemia is unknown. We found human polymorphonuclear leukocytes (PMNs) were a significant source of PAF receptor agonists after stimulation by either class of endotoxin. Production of PAF receptor agonists required that the PMN adhere to a surface, and adhesion (and therefore accumulation of PAF-like bioactivity) in response to endotoxic stimulation was delayed for several minutes. PAF-like oxidized phospholipids were found by mass spectroscopy, but biosynthetic PAF accounted for most of the phospholipid agonists arising from endotoxic stimulation. A significant portion of the PAF made by PMNs was secreted, in contrast to its near complete retention by other inflammatory cells. Endotoxic stimulation induced a respiratory burst with the production of superoxide and the formation and shedding of microparticles. Free and microparticle-bound PAF appeared in the media, and blocking microvesiculation with calpeptin blocked PAF release. The released material activated platelets, and platelets co-aggregated with endotoxin-stimulated PMNs. Adherent PMNs therefore behave differently than suspended cells and are a significant source of free PAF after endotoxin exposure. Leukocytes can couple endotoxic challenge to the widespread circulatory and inflammatory effects of endotoxin.
authors.affiliation: Department of Human Molecular Biology and Genetics, University of Utah, Salt Lake City, Utah 84112-5330, USA.
authors.forename: Junji
authors.initials: J
authors.lastname: Watanabe
authors.forename: Gopal K
authors.initials: GK
authors.lastname: Marathe
authors.forename: Paul O
authors.initials: PO
authors.lastname: Neilsen
authors.forename: Andrew S
authors.initials: AS
authors.lastname: Weyrich
authors.forename: Kathleen A
authors.initials: KA
authors.lastname: Harrison
authors.forename: Robert C
authors.initials: RC
authors.lastname: Murphy
authors.forename: Guy A
authors.initials: GA
authors.lastname: Zimmerman
authors.forename: Thomas M
authors.initials: TM
authors.lastname: McIntyre
mesh_terms: D001792:Blood Platelets / Q000378:metabolism; D002118:Calcium / Q000378:metabolism; D002448:Cell Adhesion; D002460:Cell Line; D004731:Endotoxins / Q000378:metabolism*; D006801:Humans; D007249:Inflammation; D008024:Ligands; D050356:Lipid Metabolism; D013058:Mass Spectrometry; D018613:Microscopy, Confocal; D009504:Neutrophils / Q000166:cytology*; D010100:Oxygen / Q000378:metabolism; D010972:Platelet Activating Factor / Q000378:metabolism*; D013696:Temperature; D013997:Time Factors
chemical_list: D004731:Endotoxins; D008024:Ligands; D010972:Platelet Activating Factor; D010100:Oxygen; D002118:Calcium