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Volume 341, Issue 1 p. 65-68
Research letters
Free Access

Kinetics of nitric oxide and hydrogen peroxide production and formation of peroxynitrite during the respiratory burst of human neutrophils

Maria Cecilia Carreras

Corresponding Author

Maria Cecilia Carreras

Laboratory of Oxygen Metabolism, University Hospital, University of Buenos Aires, Cordoba 2351, 1120 Buenos Aires, Argentina

Corresponding author. Fax: (54) (1) 961-6148.Search for more papers by this author
Griselda A. Pargament

Griselda A. Pargament

Laboratory of Oxygen Metabolism, University Hospital, University of Buenos Aires, Cordoba 2351, 1120 Buenos Aires, Argentina

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Sergio D. Catz

Sergio D. Catz

Laboratory of Oxygen Metabolism, University Hospital, University of Buenos Aires, Cordoba 2351, 1120 Buenos Aires, Argentina

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Juan J. Poderoso

Juan J. Poderoso

Laboratory of Oxygen Metabolism, University Hospital, University of Buenos Aires, Cordoba 2351, 1120 Buenos Aires, Argentina

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Alberto Boveris

Alberto Boveris

Laboratory of Oxygen Metabolism, University Hospital, University of Buenos Aires, Cordoba 2351, 1120 Buenos Aires, Argentina

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First published: March 14, 1994
Citations: 280

Abstract

Nitric oxide (.NO) release, oxygen uptake and hydrogen peroxide (H2O2) production elicited by increasing phorbol 12-myristate 13-acetate (PMA) concentrations were measured in human neutrophils. Half-maximal activities were sequentially elicited at about 0.0001–0.001 μg urn:x-wiley:00145793:media:feb20014579394802416:feb20014579394802416-math-si1 (.NO) and 0.001-0.01 μg urn:x-wiley:00145793:media:feb20014579394802416:feb20014579394802416-math-si2 (H2O2). At saturated PMA concentrations, .NO production, oxygen uptake and H2O2 release were 0.56 ± 0.04, 3.32 ± 0.52 and 1.19±0.17 nmol · min−1 · 106 cells−1. .NO production accounts for about 30% of the total oxygen uptake. Luminol-dependent chemiluminescence, reported to detect NO reactions in other inflammatory cells, was also half-maximally activated at about 0.001-0.01 μg urn:x-wiley:00145793:media:feb20014579394802416:feb20014579394802416-math-si3. Preincubation with N G-monomethyl-l-arginine (l-NMMA) decreased O2 uptake and .NO release but increased H2O2 production, while superoxide dismutase (SOD) increased .NO detection by 30%. Chemiluminescence was also reduced by preincubation with l-NMMA and/or SOD. The results indicate that .NO release is part of the integrated response of stimulated human neutrophils and that, in these cells, kinetics of ″NO and O2 .− release favour the formation of other oxidants like peroxynitrite.