title: Involvement of platelet-activating factor in endotoxin-induced priming of rabbit polymorphonuclear leukocytes.
abstract: Endotoxin primes polymorphonuclear leukocytes (PMNs) for increased superoxide anion (O2-) generation in response to the chemotactic peptide, formyl-methionyl-leucyl-phenylalanine (fMLP). We have investigated the role of platelet-activating factor (PAF) in the priming of rabbit PMNs by endotoxin. In non-primed PMNs, fMLP stimulated PAF generation, but none was detected in the supernatant, whereas in primed PMNs PAF generation increased 17-fold and a large proportion of the total PAF formed was released. There was a close relationship between the concentration-response curve for increases in PAF synthesis and that for enhanced O2- generation. The possibility that PAF was causally linked to enhanced O2- generation in primed PMNs was investigated using compounds previously reported to reduce PAF generation, namely the serine protease inhibitor, tosyl-phenylalanine chloromethylketone, and the phospholipase A2 inhibitor, bromophenacylbromide. Both compounds reduced PAF generation and release in endotoxin-primed PMNs and prevented the enhancement of O2- generation. We examined the possibility that PAF acted in an autocrine fashion to enhance O2- generation following its release from fMLP-stimulated PMNs. However, rabbit PMNs, unlike human PMNs, were not primed by exogenous PAF (0.1-1000 nM) over a concentration range extending beyond that covering the concentrations of PAF determined in the supernatants of fMLP-stimulated, endotoxin-primed PMNs. We conclude that PAF release is unlikely to play a role in priming induced by endotoxin in rabbit PMNs, but our data are not inconsistent with a role for cell-associated PAF in this phenomenon. Furthermore, the release of a large amount (80%) of the total PAF from primed, but not non-primed PMNs suggests a cell-cell mediator role for PAF only in primed PMNs.
authors.affiliation: Department of Physiology, University of Melbourne, Parkville, Vic., Australia.
authors.forename: A G
authors.initials: AG
authors.lastname: Stewart
authors.forename: T
authors.initials: T
authors.lastname: Harris
mesh_terms: D000098:Acetophenones / Q000494:pharmacology; D000818:Animals; D000001:Calcimycin / Q000494:pharmacology; D004731:Endotoxins / Q000378:metabolism*; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000494:pharmacology; D009504:Neutrophils / Q000187:drug effects / Q000378:metabolism*; D010972:Platelet Activating Factor / Q000378:metabolism*; D011817:Rabbits; D015398:Signal Transduction; D013481:Superoxides / Q000032:analysis / Q000378:metabolism*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014108:Tosylphenylalanyl Chloromethyl Ketone / Q000494:pharmacology
chemical_list: D000098:Acetophenones; D004731:Endotoxins; D010972:Platelet Activating Factor; D013481:Superoxides; D000001:Calcimycin; D014108:Tosylphenylalanyl Chloromethyl Ketone; D009240:N-Formylmethionine Leucyl-Phenylalanine; C013190:phenacyl bromide; D013755:Tetradecanoylphorbol Acetate