This paper is only available as a PDF. To read, Please Download here.
Abstract
The production of nitric oxide (NO) may play an important role in functional responses
of the human polymorphonuclear neutrophil granulocytes (PMNs). Others have described
the presence of both an inducible, Ca2+-independent and a constitutionally expressed, Ca2+-dependent nitric oxide synthase (NOS) in human PMNs. However, the conditions for
production and release of NO in human PMNs are still largely unknown. We assessed
mechanisms for activation of NO release from human PMNs and particularly the dependence
on extracellular and intracellular Ca2+. We addressed this question by applying a variety of agonists with known and differing
mechanisms of activation in PMNs and measuring the released NO by two highly sensitive
and specific real-time methods for detection of NO, the oxidation of oxyhemoglobin
to methemoglobin and an electrochemical method. We found that human PMNs activated
with the surface receptor-dependent agonist, N-formyl-methionyl-leucyl-phenylalanine (fMLP); the calcium lonophore, A23187; or the
direct stimulator of protein kinase C, phorbol myristate acetate (PMA), produced NO
which was inhibited by a specific NOS inhibitor, NG-monomethyl-l-arginine. The NO production induced by fMLP or A23187 was dependent on the presence
of extracellular Ca2+, but this was not the case for PMA. The stimulatory effect of fMLP was almost completely
inhibited by Bordetella pertussis toxin. These results indicate an NOS activity in purified human PMNs in vitro, and
the transduction mechanisms for the agonists used show strong similarity with previously
known pathways for other neutrophil functions.
Abbreviations:
BAPTA (1,2-bis(2-Aminophenoxy)-ethane-N,N,N′,N′-tetraacetic acid), BAPTAAM (BAPTA-aetoxymethylester), cNOS (constitutive NOS), EGTA (ethyleneglycol-bis-(β-aminoethylether)-N,N,N′,N′-tetraacetic acid), fMLP (N-formyl-methionyl-leucyl-phenylalanine), HBSS (Hanks balanced salt solution), iNOS (inducible NOS), L-NMMA (NG-monomethyl-l-arginine), NADPH (the reduced form of nicotinamide-adenine dinucleotide phosphate), NOS (nitric oxide synthase), ·O2− (superoxide anion), PMA (phorbol myristate acetate), PMN (polymorphonuclear neutrophil granulocyte), PT (pertussis toxin), SNAP (S-nitroso-acetylpenicillamine), SOD (superoxide dismutase)To read this article in full you will need to make a payment
Purchase one-time access:
Academic & Personal: 24 hour online accessCorporate R&D Professionals: 24 hour online accessOne-time access price info
- For academic or personal research use, select 'Academic and Personal'
- For corporate R&D use, select 'Corporate R&D Professionals'
Subscribe:
Subscribe to Translational ResearchAlready a print subscriber? Claim online access
Already an online subscriber? Sign in
Register: Create an account
Institutional Access: Sign in to ScienceDirect
References
- Nitric oxide as a mediator of inflammation.Int J Immunopathol Pharmacol. 1993; 6: 67-75
- Nitric oxide synthases in mammals.Biochem J. 1994; 298: 249-258
- Regulation of biosynthesis of nitric oxide.J Biol Chem. 1994; 269: 13725-13728
- Isoforms of nitric oxide synthase.Biochem Pharmacol. 1991; 42: 1849-1857
- Ca2+/calmodulin-regulated nitric oxide synthases.Cell Calcium. 1992; 13: 427-434
- Molecular cloning and functional expression of an inducible nitric oxide synthase from a murine macrophages cell line.J Biol Chem. 1992; 267: 6370-6374
- Induction of nitric oxide synthase in rat peritoneal neutrophils and its inhibition by dexamethason.Eur J Immunol. 1991; 21: 2523-2527
- Human polymorphonuclear leukocytes lack detectable nitric oxide synthase activity.J Immunol. 1994; 153: 1825-1834
- Co-purification of 130 kd nitric oxide synthase and a 22 kd link protein from human neutrophils.Biochem Biophys Res Commun. 1992; 189: 558-564
- Identification of the NO synthase isoforms expressed in human neutrophil granulocytes, megakaryocytes and platelets.Thromb Haemost. 1997; 77: 163-167
- Bacterial infection induces nitric oxide synthase in human neutrophils.J Clin Invest. 1997; 99: 110-116
- Kinetics of nitric oxide and hydrogen peroxide production and formation of peroxynitrite during the respiratory burst of human neutrophils.FEBS Lett. 1994; 341: 65-68
- Generation of nitric oxide by human neutrophils.Biochem Biophys Res Commun. 1989; 160: 813-819
- Nitric oxide production by activated human neutrophils exposed to sodium azide and hydroxylamine: The role of oxygen radicals.Biochem Biophys Res Commun. 1994; 199: 1245-1249
- Endogenous nitric oxide: Physiology, pathology and relevance.Eur J Clin Invest. 1991; 21: 361-374
- Nitric oxide, an endothelial cell relaxation factor, inhibits neutrophil superoxide anion production via a direct action on the NADPH oxidase.J Clin Invest. 1992; 90: 1116-1121
- Nitric oxide inactivates NADPH oxidase in pig neutrophils by inhibiting its assembling process.J Biol Chem. 1997; 272: 32773-32778
- Signal transduction mechanisms in phagocyte priming and agonist responses.Curr Opin Hematol. 1993; 1: 106-112
- Neutrophil signal transduction: Calcium, kinases, and fusion.J Lab Clin Med. 1992; 120: 527-612
- Chemoattractant receptors.Curr Opin Hematol. 1994; 1: 40-44
- Protein kinase C and the activation of the human neutrophil NADPH-oxidase.Blood. 1987; 69: 711-720
- Measurement of methemoglobin formation from oxyhemoglobin: A real-time, continuous assay of nitric oxide release by human polymorphonuclear leukocytes.J Immunol Methods. 1995; 184: 53-62
- The biochemical pathways of nitric oxide formation from nitrovasodilators: Appropriate choice of exogenous NO donors and aspects of preparation and handling of aqueous NO solutions.J Cardiovasc Pharmacol. 1991; 17: S25-S33
- Nitric oxide production in polymorphonuclear granulocytes.Endothelium. 1993; 1 (suppl): 31
- The L-arginine: nitric oxide pathway.Acta Physiol Scand. 1992; 145: 201-227
- Correlation between nitric oxide formation during degradation of organic nitrates and activation of guanylate cyclase.Eur J Pharmacol. 1987; 139: 19-30
- Selective and sensitive electrochemical measurement of nitric oxide in aqueous solution: Discussion and new results.J Electroanal Chem. 1995; 392: 85-89
- Direct measurement of nitric oxide production in platelets: Relationship with cytosolic Ca2+ concentration.Biochem Biophys Res Commun. 1995; 215: 842-848
- Lucigenin chemiluminescence in the assessment of neutrophil superoxide production.J Immunol Methods. 1987; 97: 209-213
- A new generation of Ca2+ indicators with greatly improved fluorescence properties.J Biol Chem. 1985; 260: 3440-3450
- Thrombin and ionomycin can raise platelet cytosolic Ca2+ to micromolar levels by discharge of internal Ca2+ stores: Studies using Fura-2.Biochem Biophys Res Commun. 1986; 139: 308-314
- Tyrosine phosphorylation in activated human neutrophils: Comparison of the effects of different classes of agonists and identification of the signaling pathways involved.J Immunol. 1994; 153: 353-363
- The interactive effects of fluoride and N-formyl-Lmethionyl-L-leucyl-L-phenylalanine on superoxide production and cAMP levels in human neutrophils.Can J Biochem Cell Biol. 1982; 61: 569-578
Article info
Publication history
Accepted:
March 13,
1998
Received in revised form:
March 5,
1998
Received:
October 2,
1997
Footnotes
☆Supported by the Swedish Medical Research Council (Grant 19X05991), the Funds of the Karolinska Institute, the Swedish Medical Association, the Swedish Heart and Lung Association, King Gustaf V Fund, Tore Nilson's Fund, the Funds of Borje Dahlin, Inga-Britt and Arne Lundberg, and the Swedish Association Against Rheumatism
Identification
Copyright
© 1998 Published by Elsevier Inc.
