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Original article| Volume 132, ISSUE 1, P54-60, July 1998

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Stimulus-dependent transduction mechanisms for nitric oxide release in human polymorphonuclear neutrophil leukocytes

  • Gerd Lärfars
    Correspondence
    Reprint requests: Gerd Lärfars, MD, Department of Hematology, M54, Huddinge University Hospital, S-141 86 Huddinge, Sweden.
    Affiliations
    Department of Hematology and the Laboratory for Inflammation and Hematology Research, the Karolinska Institute at Huddinge University Hospital, Huddinge, Sweden
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  • Hans Gyllenhammar
    Affiliations
    Department of Hematology and the Laboratory for Inflammation and Hematology Research, the Karolinska Institute at Huddinge University Hospital, Huddinge, Sweden
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      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)
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      References

        • Gorbunov N
        • Esposito E
        Nitric oxide as a mediator of inflammation.
        Int J Immunopathol Pharmacol. 1993; 6: 67-75
        • Knowles R
        • Moncada S
        Nitric oxide synthases in mammals.
        Biochem J. 1994; 298: 249-258
        • Nathan C
        • Xie Q
        Regulation of biosynthesis of nitric oxide.
        J Biol Chem. 1994; 269: 13725-13728
        • Porstermann U
        • Schmidt HHW
        • Pollock JS
        • Sheng H
        • Mitchell JA
        • Warner TD
        • et al.
        Isoforms of nitric oxide synthase.
        Biochem Pharmacol. 1991; 42: 1849-1857
        • Schmidt HHW
        • Pollock JS
        • Nakane M
        • Forstermann U
        • Murad F
        Ca2+/calmodulin-regulated nitric oxide synthases.
        Cell Calcium. 1992; 13: 427-434
        • Lyons CR
        • Orloff GJ
        • Cunningham JM
        Molecular cloning and functional expression of an inducible nitric oxide synthase from a murine macrophages cell line.
        J Biol Chem. 1992; 267: 6370-6374
        • McCall TB
        • Palmer RMJ
        • Moncada S
        Induction of nitric oxide synthase in rat peritoneal neutrophils and its inhibition by dexamethason.
        Eur J Immunol. 1991; 21: 2523-2527
        • Yan L
        • Vandivier RW
        • Suffredini AF
        • Danner RL
        Human polymorphonuclear leukocytes lack detectable nitric oxide synthase activity.
        J Immunol. 1994; 153: 1825-1834
        • Bryant JL
        • Mehta P
        • Vanderporten A
        • Mehta JL
        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
        • Wallerath T
        • Gath I
        • Aulitzky WE
        • Pollock JS
        • Kleinert H
        • Förstermann U
        Identification of the NO synthase isoforms expressed in human neutrophil granulocytes, megakaryocytes and platelets.
        Thromb Haemost. 1997; 77: 163-167
        • Wheeler A
        • Shannon D
        • Smith Guillermo GC
        • Nathan CF
        • Weiss RM
        • William C
        Bacterial infection induces nitric oxide synthase in human neutrophils.
        J Clin Invest. 1997; 99: 110-116
        • Carreras MC
        • Pargament GA
        • Catz SD
        • Poderoso JJ
        • Boveris A
        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
        • Wright CD
        • Mulsch A
        • Busse R
        • Osswald H
        Generation of nitric oxide by human neutrophils.
        Biochem Biophys Res Commun. 1989; 160: 813-819
        • Markert M
        • Carnal B
        • Mauel J
        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
        • Moneada S
        • Higgs A
        Endogenous nitric oxide: Physiology, pathology and relevance.
        Eur J Clin Invest. 1991; 21: 361-374
        • Clancy RM
        • Leszcynska-Piziak J
        • Abramson SB
        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
        • Fuji H
        • Ichimori K
        • Hosbiai K
        • Nakazawa H
        Nitric oxide inactivates NADPH oxidase in pig neutrophils by inhibiting its assembling process.
        J Biol Chem. 1997; 272: 32773-32778
        • Lew DP
        • Krause KH
        Signal transduction mechanisms in phagocyte priming and agonist responses.
        Curr Opin Hematol. 1993; 1: 106-112
        • Smolen JE
        Neutrophil signal transduction: Calcium, kinases, and fusion.
        J Lab Clin Med. 1992; 120: 527-612
        • Perez HD
        Chemoattractant receptors.
        Curr Opin Hematol. 1994; 1: 40-44
        • Tauber AI
        Protein kinase C and the activation of the human neutrophil NADPH-oxidase.
        Blood. 1987; 69: 711-720
        • Larfars G
        • Gyllenhammar H
        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
        • Feelisch M
        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
        • Lirfars G
        • Gyllenhammar H
        Nitric oxide production in polymorphonuclear granulocytes.
        Endothelium. 1993; 1 (suppl): 31
        • Moncada S
        The L-arginine: nitric oxide pathway.
        Acta Physiol Scand. 1992; 145: 201-227
        • Feelisch M
        • Noack E
        Correlation between nitric oxide formation during degradation of organic nitrates and activation of guanylate cyclase.
        Eur J Pharmacol. 1987; 139: 19-30
        • Lantoine F
        • Trevin S
        • Bedioui F
        • Devynck J
        Selective and sensitive electrochemical measurement of nitric oxide in aqueous solution: Discussion and new results.
        J Electroanal Chem. 1995; 392: 85-89
        • Lantoine F
        • Brunet A
        • Bedioui F
        • Devynck J
        • Devynck MA
        Direct measurement of nitric oxide production in platelets: Relationship with cytosolic Ca2+ concentration.
        Biochem Biophys Res Commun. 1995; 215: 842-848
        • Gyllenhammar H
        Lucigenin chemiluminescence in the assessment of neutrophil superoxide production.
        J Immunol Methods. 1987; 97: 209-213
        • Grynkiewicz G
        • Poenie M
        • Tsien RT
        A new generation of Ca2+ indicators with greatly improved fluorescence properties.
        J Biol Chem. 1985; 260: 3440-3450
        • Pollock WK
        • Rink TJ
        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
        • Rollet E
        • Caon AC
        • Roberge CJ
        • Liao NW
        • Malawista SE
        • McColl SR
        • et al.
        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
        • Wong K
        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