Lucigenin chemiluminescence in the assessment of neutrophil superoxide production.
Lucigenin-amplified chemiluminescence (LUCL) was induced by .O2- (from the xanthine/xanthine-oxidase reaction) but not, as with luminol-augmented CL (LCL), by myeloperoxidase (MPO) and only weakly by H2O2 or the H2O2-MPO-Cl- system. Neutrophil LUCL induced by fMet-Leu-Phe (fMLP) was dose-dependently inhibited by scavengers of .O2- but not by NaN3, catalase, mannitol or taurine. Response patterns of several soluble stimuli (leukotriene B4 (LTB4), platelet-activating factor (PAF), fMLP, A23187 and phorbol-myristate-acetate (PMA)) were assessed with LUCL. PMA-induced LUCL was protracted, A23187 showed intermediary kinetics whereas fMLP and PAF both showed rapid peak responses at 30-50 s. However, LTB4 was the most rapid initiator of LUCL (7-12 s). The kinetics of fMLP- or PMA-induced neutrophil .O2- production, assessed with the cytochrome C reduction method, correlated well with LUCL. Thus it is suggested that LUCL provides a specific method for studying the kinetic properties of neutrophil .O2- production where stimulus-specific response patterns can be distinguished.
D000166:Acridines; D005609:Free Radicals; D006801:Humans; D007975:Leukotriene B4 / Q000494:pharmacology; D008163:Luminescent Measurements; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000494:pharmacology; D009504:Neutrophils / Q000378:metabolism*; D009195:Peroxidase / Q000494:pharmacology; D013481:Superoxides / Q000378:metabolism*
D000166:Acridines; D005609:Free Radicals; D013481:Superoxides; D007975:Leukotriene B4; C033472:10,10'-dimethyl-9,9'-biacridinium; D009240:N-Formylmethionine Leucyl-Phenylalanine; D009195:Peroxidase