Oxidant-scavenging activities of ampicillin and sulbactam and their effects on neutrophil functions.
Luminol-enhanced luminescence is a method used to measure formation of reactive oxygen intermediates important in the ability of neutrophils to kill microbes. Several studies have demonstrated that under some conditions of incubation, ampicillin can inhibit neutrophil-derived luminol-enhanced luminescence. We evaluated the mechanism(s) by which ampicillin inhibited the luminescent response of stimulated neutrophils. We also investigated sulbactam, a beta-lactamase inhibitor which has been given in combination with ampicillin and other beta-lactam antibiotics to increase their spectra, for possible similar effects. Both ampicillin and sulbactam attenuated luminol-enhanced luminescence by approximately 40%. Superoxide production was not prevented by added ampicillin, nor was superoxide scavenged by it. Myeloperoxidase reacts with H2O2 and Cl- to generate OCl-, which is believed to be the oxidizer of luminol that is primarily responsible for enhancement of neutrophil-derived luminescence. Hydroxyl radicals (HO.), which may also oxidize luminol, resulting in luminescence, can be formed from O2- and H2O2 via either myeloperoxidase-dependent (involving intermediate OCl-) or myeloperoxidase-independent (through a metal ion catalyst) reactions. Ampicillin scavenged H2O2 and OCl- and prevented 95% of Fenton reaction-generated HO. from reacting with 5,5-dimethyl-1-pyrroline-N-oxide. Sulbactam was found to scavenge OCl- and HO., but less avidly than ampicillin did. Neither ampicillin nor sulbactam inhibited myeloperoxidase activity. Sublethal concentrations of sulbactam had no significant effect on neutrophil killing of Staphylococcus aureus and Escherichia coli. Our results demonstrate a mechanism(s) by which ampicillin inhibits luminol-enhanced luminescence from stimulated neutrophils, namely, through scavenging of the oxidant(s) primarily responsible for the generation of luminescence.
Department of Medicine, University of North Carolina, Chapel Hill 27599-7030.
M R
MR
Gunther
J
J
Mao
M S
MS
Cohen
D000667:Ampicillin / Q000378:metabolism / Q000494:pharmacology*; D001770:Blood Bactericidal Activity / Q000187:drug effects; D016166:Free Radical Scavengers; D006801:Humans; D008163:Luminescent Measurements; D009504:Neutrophils / Q000187:drug effects* / Q000378:metabolism / Q000382:microbiology; D016877:Oxidants / Q000378:metabolism / Q000494:pharmacology*; D009195:Peroxidase / Q000378:metabolism; D012973:Sodium Hypochlorite / Q000378:metabolism; D013407:Sulbactam / Q000378:metabolism / Q000494:pharmacology*; D013654:Taurine / Q000031:analogs & derivatives / Q000378:metabolism; D015054:Zymosan / Q000494:pharmacology
D016166:Free Radical Scavengers; D016877:Oxidants; D013654:Taurine; C043410:N-chlorotaurine; D000667:Ampicillin; D015054:Zymosan; D012973:Sodium Hypochlorite; D009195:Peroxidase; D013407:Sulbactam