Naunyn-Schmiedeberg's Archives of Pharmacology

, Volume 343, Issue 4, pp 370–376 | Cite as

Differential inhibition and potentiation of chemoattractant-induced superoxide formation in human neutrophils by the cell-permeant analogue of cyclic GMP, N2,2′-O-dibutyryl guanosine 3′:5′-cyclic monophosphate

  • Jürgen Ervens
  • Günter Schultz
  • Roland Seifert


Human neutrophils possess a superoxide (O inf2 sup− )-forming NADPH oxidase which is activated by the chemoattractants, N-formyl-l-methionyl-l-leucyl-l,-phenylalanine (fMet-Leu-Phe), complement C5a, platelet-activating factor and leukotriene B4. We studied the roles of cAMP and cGMP in the regulation of O inf2 sup− formation using the cell-permeant analogues of cyclic nucleotides, N6,2′-O-dibutyryl adenosine 3′:5′-cyclic monophosphate (Bt2cAMP) and N2,2′-O-dibutyryl guanosine 3′:5′-cyclic monophosphate (Bt2cGMP). Bt2cAMP inhibited O inf2 sup− formation induced by these chemoattractants to similar extents. Bt2cGMP as low as 10 μmol/l significantly inhibited O inf2 sup− formation induced by fMet-Leu-Phe at a submaximally effective concentration (50 nmol/1), and Bt2cGMP was more effective in diminishing O inf2 sup− formation than Bt2cAMP. In contrast, Bt2cGMP did not affect O inf2 sup− formation induced by fMet-Leu-Phe at a maximally effective concentration (1 μmol/l). Bt2cGMP (0.1 and 1 mmol/l) enhanced O inf2 sup− formation induced by 0.1 μmol/l C5a by 23% and 49%, respectively, and Bt2cGMP antagonized inhibition of O inf2 sup− formation caused by Bt2cAMP. Bt2cGMP inhibited platelet-activating factor-induced O inf2 sup− formation to a lesser extent than Bt2cAMP and had no effect on that induced by leukotriene B4. Bt2cAMP and Bt2cGMP had no effect on O inf2 sup− formation induced by NaF, γ-hexachlorocyclohexane, phorbol myristate acetate, A 23187 and arachidonic acid. Our data suggest that: 1. Bt2cAMP generally inhibits chemoattractant-stimulated O inf2 sup− formation. 2. Bt2cGMP inhibits fMet-Leu-Phe- and platelet-activating factor-stimulated O inf2 sup− formation but potentiates C5a-induced O inf2 sup− formation. 3. The lack of effect of cyclic nucleotides on O inf2 sup− formation induced by agents other than receptor agonists indicates that cAMP and cGMP modulate early steps of the signal transduction processes initiated by chemoattractants.

Key words

N6,2′-O-dibutyryl adenosine 3′:5′-cyclic monophosphate N2,2′-O-dibutyryl guanosine 3′:5′ 


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  1. Beavo JA (1988) Multiple isozymes of cyclic nucleotide phosphodiesterase. Adv Second Messenger Phosphoprotein Res 22:1–38PubMedGoogle Scholar
  2. Bender JG, van Epps DE, Chenoweth DE (1987) Independent regulation of human neutrophil chemotactic receptors after activation. J Immunol 139:3028–3033PubMedGoogle Scholar
  3. Böhme E, Graf H, Schultz G (1978) Effects of sodium nitroprusside and other smooth muscle relaxants on cyclic GMP formation in smooth muscle and platelets. Adv Cycl Nucl Res 9:131–143Google Scholar
  4. Boss GR (1989) cGMP-induced differentiation of the promyelocytic cell line HL-60. Proc Natl Acad Sci USA 86:7174–7178CrossRefGoogle Scholar
  5. Burde R, Seifert R, Buschauer A, Schultz G (1989) Histamine inhibits activation of human neutrophils and HL-60 leukemic cells via H2-receptors. Naunyn-Schmiedeberg's Arch Pharmacol 340:671–678CrossRefGoogle Scholar
  6. Ignarro LJ, George WJ (1974) Hormonal control of lysosomal enzyme release from human neutrophils: Elevation of cyclic nucleotide levels by autonomic neurohormones. Proc Natl Acad Sci USA 71:2027–2031CrossRefGoogle Scholar
  7. Jesaitis AJ, Tolley JO, Allen RA (1986) Receptor-cytoskeleton interactions and membrane traffic may regulate chemoattractant-induced superoxide production in human granulocytes. J Biol Chem 261:13662–13669PubMedGoogle Scholar
  8. Kaplan SS, Billiar T, Zdziarski UE, Simmons RL, Basford RE (1989) Inhibition of chemotaxis with NG-monomethyl-l-arginine: A role for cyclic GMP. Blood 74:1885–1887PubMedGoogle Scholar
  9. Kim U-H, Kim JW, Rhee SG (1989) Phosphorylation of phospholipase C-y by cAMP-dependent protein kinase. J Biol Chem 264:20167–20170PubMedGoogle Scholar
  10. Kramer IM, van der Bend RL, Verhoeven AJ, Roos D (1988) The 47-kDa protein involved in the NADPH:O2 oxidoreductase activity of human neutrophils is phosphorylated by cyclic AMP-dependent protein kinase without induction of a respiratory burst. Biochim Biophys Acta 971:189–196PubMedGoogle Scholar
  11. Kuo JF, Shoji M, Kuo W-N (1978) Molecular and physiopathologic aspects of mammalian cyclic GMP-dependent protein kinase. Annu Rev Pharmacol Toxicol 18:341–355CrossRefGoogle Scholar
  12. Lad PM, Glovsky MM, Richards JH, Smiley PA, Backstrom B (1985) Regulation of human neutrophil guanylate cyclase by metal ions, free radicals and the muscarinic cholinergic receptor. Mol Immunol 22:731–739CrossRefGoogle Scholar
  13. Lefkowitz RJ, Caron MC (1986) Regulation of adrenergic receptor function by phosphorylation. Curr Top Cell Regul 28:209–231CrossRefGoogle Scholar
  14. Malech HL, Gallin MD (1987) Neutrophils in human diseases. N Engl J Med 317:687–694CrossRefGoogle Scholar
  15. McPhail LC, Clayton CC, Snyderman R (1984) The NADPH oxidase of human polymorphonuclear leucocytes. Evidence for regulation by multiple signals. J Biol Chem 259:5768–5775PubMedGoogle Scholar
  16. Misaki N, Imaizumi T, Watanabe Y (1989) Cyclic AMP-dependent protein kinase interferes with GTPTS stimulated IP3 formation in differentiated HL-60 cell membranes. Life Sci 45:1671–1678CrossRefGoogle Scholar
  17. Moncada S, Palmer RMJ, Higgs EA (1989) Biosynthesis of nitric oxide from l-arginine. A pathway for the regulation of cell function and communication. Biochem Pharmacol 38:1709–1715CrossRefGoogle Scholar
  18. Mueller H, Sklar LA (1989) Coupling of antagonistic signalling pathways in modulation of neutrophil function. J Cell Biochem 40:287–294CrossRefGoogle Scholar
  19. Naor Z (1990) Cyclic GMP stimulates inositol phosphate production in cultured pituitary cells: Possible implication to signal transduction. Biochem Biophys Res Commun 167:982–992CrossRefGoogle Scholar
  20. Pryzwansky KB, Wyatt TA, Nichols H, Lincoln TM (1990) Compartmentalization of cyclic GMP-dependent protein kinase in formyl-peptide stimulated neutrophils. Blood 76:612–618PubMedGoogle Scholar
  21. Rossi F (1986) The O2-forming NADPH oxidase of the phagocytes: nature, mechanisms of activation and function. Biochim Biophys Acta 853:65–89CrossRefGoogle Scholar
  22. Schmidt HHHW, Seifert R, Böhme E (1989) Formation and release of nitric oxide from human neutrophils and HL-60 cells induced by a chemotactic peptide, platelet activating factor and leukotriene B4. FEBS Lett 244:357–360CrossRefGoogle Scholar
  23. Schröder H, Ney P, Woditsch I, Schrör K (1990) Cyclic GMP mediates SIN-1-induced inhibition of human polymorphonuclear leukocytes. Eur J Pharmacol 182:211–218CrossRefGoogle Scholar
  24. Schultz K-D, Böhme E, Kreye VAW, Schultz G (1979) Relaxation of hormonally stimulated smooth muscular tissues by the 8-bromo derivative of cyclic GMP. Naunyn-Schmiedeberg's Arch Pharmacol 306:1–9CrossRefGoogle Scholar
  25. Seifert R, Schultz G (1991) The superoxide-forming NADPH oxidase of phagocytes: An enzyme system regulated by multiple mechanisms. Rev Physiol Biochem Pharmacol (in press)Google Scholar
  26. Seifert R, Burde R, Schultz G (1989a) Activation of NADPH oxidase by purine and pyrimidine nucleotides involves G proteins and is potentiated by chemotactic peptides. Biochem J 259:813–819CrossRefGoogle Scholar
  27. Seifert R, Burde R, Schultz G (1989b) Lack of effect of opioid peptides, morphine and naloxone on superoxide formation in human neutrophils and HL-60 leukemic cells. Naunyn-Schmiedeberg's Arch Pharmacol 340:101–106CrossRefGoogle Scholar
  28. Seifert R, Wenzel K, Eckstein F, Schultz G (1989c) Purine and pyrimidine nucleotides potentiate activation of NADPH oxidase and degranulation by chemotactic peptides and induce aggregation of human neutrophils via G proteins. Eur J Biochem 181:277–285CrossRefGoogle Scholar
  29. Seifert R, Jungblut P, Schultz G (1989d) Differential expression of cytosolic activation factors for NADPH oxidase in HL-60 leukemic cells. Biochem Biophys Res Commun 161:1109–1117CrossRefGoogle Scholar
  30. Tremblay J, Gerzer R, Hamet P (1988) Cyclic GMP in cell function. Adv Second Messenger Phosphoprotein Res 22:319–383PubMedGoogle Scholar
  31. Waldman SA, Murad F (1987) Cyclic GMP synthesis and function. Pharmacol Rev 39:163–196PubMedGoogle Scholar
  32. Walter U (1989) Physiological role of cGMP and cGMP-dependent protein kinase in the cardiovascular system. Rev Physiol Biochem Pharmacol 113:41–88CrossRefGoogle Scholar
  33. Wright CD, Mülsch A, Busse R, Osswald H (1989) Generation of nitric oxide by human neutrophils. Biochem Biophys Res Commun 160:813–819CrossRefGoogle Scholar
  34. Wymann MP, von Tscharner V, Daranleau DA, Baggiolini M (1987) The onset of the respiratory burst in human neutrophils. Real-time studies of H2O2 formation reveal a rapid agonist-induced transduction process. J Biol Chem 262:12048–12053PubMedGoogle Scholar

Copyright information

© Springer-Verlag 1991

Authors and Affiliations

  • Jürgen Ervens
    • 1
  • Günter Schultz
    • 1
  • Roland Seifert
    • 1
  1. 1.Institut für PharmakologieFreie Universität BerlinBerlin 33Germany

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