Mechanisms for luminol-augmented chemiluminescence from neutrophils induced by leukotriene B4 and N-formyl-methionyl-leucyl-phenylalanine.
Neutrophils stimulated with formyl-methionyl-leucyl-phenylalanine (fMLP) or leukotriene B4 (LTB4) generated kinetically distinctive luminol augmented chemiluminescence (LCL). Inhibitors of .O2- [superoxide-dismutase (SOD) or tiron], H2O2 (catalase), myeloperoxidase, MPO, (NaN3), HOCl (taurine) and .OH (mannitol) hampered LCL dose-dependently with similar characteristics for both stimuli. In cell free systems it was found that .O2- (generated in the xanthine/xanthine-oxidase reaction) or H2O2 produced LCL. Superoxide dismutase inhibited .O2- -induced LCL dose dependently. The MPO + H2O2 system, which generated more pronounced LCL than either component alone, was inhibited by catalase and taurine but not by SOD. When neutrophils, treated with luminol, but where extracellular luminol had been removed, were stimulated with fMLP or LTB4, they produced less than 2% of the LCL where luminol was present in the medium. When neutrophil LCL and superoxide formation by the cytochrome C method were assessed in parallel experiments, in all instances the peak LCL response coincided with the linear phase in that response. Thus, LCL, induced by LTB4 and the corresponding fMLP peak, are extracellular events with similar chemical backgrounds, closely related to generation of reactive oxygen species. Consequently, the kinetical differences in LCL between fMLP and LTB4 suggest that LTB4, by yet unknown mechanisms, activates the NADPH oxidase more rapidly than fMLP.
Gyllenhammar
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D004357:Drug Synergism; D006801:Humans; D066298:In Vitro Techniques; D007975:Leukotriene B4 / Q000494:pharmacology*; D049449:Luminescence; D008165:Luminol / Q000494:pharmacology*; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000494:pharmacology*; D009504:Neutrophils / Q000187:drug effects / Q000502:physiology*; D011724:Pyridazines / Q000494:pharmacology*; D013481:Superoxides / Q000097:blood*
D011724:Pyridazines; D013481:Superoxides; D007975:Leukotriene B4; D009240:N-Formylmethionine Leucyl-Phenylalanine; D008165:Luminol