title: Stimulation of human neutrophil oxidative metabolism by nonopsonized Neisseria gonorrhoeae.
abstract: Nonopsonized gonococci possessing opacity-associated (Opa; previously PII) outer membrane proteins stimulate neutrophils to undergo a vigorous oxidative response when measured by luminol-dependent chemiluminescence (LDCL). In these studies, we characterized the mechanism of this stimulation. No gonococci that we tested induced measurable release of neutrophil superoxide anion (O2-) or hydrogen peroxide (H2O2) as measured by reduction of cytochrome c or the oxidation of scopoletin, respectively. Neutrophils pretreated with gonococci and then exposed to phorbol myristate acetate, the chemotactic peptide formylmethionylleucylphenylalanine, or opsonized zymosan released levels of neutrophil O2- and H2O2 comparable to controls, indicating that gonococci were not preventing or inhibiting neutrophil O2- or H2O2 release. To ascertain a possible explanation for these seemingly contradictory observations (i.e., induction of LDCL, but no release of O2- or H2O2), we further characterized the ability of Opa+ gonococci to stimulate LDCL. By using 1 mM azide and 4 U of horseradish peroxidase to monitor extracellular LDCL selectively and 2,000 U of catalase to monitor intracellular LDCL selectively, we determined that greater than 80% of total gonococcus-induced neutrophil LDCL occurred intracellularly. In addition, neutrophils stimulated with Opa+ gonococci showed a marked increase in O2 uptake and hexose monophosphate shunt activity. We conclude that Neisseria gonorrhoeae induces neutrophil oxidative metabolism without causing release of detectable amounts of reactive oxygen intermediates into the surrounding milieu. The gonococcus apparently directs oxidase assembly and activity to the phagolysosomal membrane. This could be a mechanism by which extracellular gonococci persist for extended periods in vivo in the presence of high concentrations of neutrophils.
authors.affiliation: Department of Microbiology and Immunology, Hahnemann University School of Medicine, Philadelphia, Pennsylvania 19102-1192.
authors.forename: F L
authors.initials: FL
authors.lastname: Naids
authors.forename: R F
authors.initials: RF
authors.lastname: Rest
mesh_terms: D000942:Antigens, Bacterial / Q000633:toxicity*; D001425:Bacterial Outer Membrane Proteins / Q000633:toxicity*; D006801:Humans; D066298:In Vitro Techniques; D008163:Luminescent Measurements; D009344:Neisseria gonorrhoeae / Q000472:pathogenicity / Q000502:physiology*; D009504:Neutrophils / Q000378:metabolism*; D010084:Oxidation-Reduction; D010101:Oxygen Consumption; D010427:Pentose Phosphate Pathway; D013481:Superoxides / Q000378:metabolism
chemical_list: D000942:Antigens, Bacterial; D001425:Bacterial Outer Membrane Proteins; C050308:opacity proteins; D013481:Superoxides