Skip to main content
Log in

Effects on extra- and intracellularly localized, chemoattractant-induced, oxygen radical production in neutrophils following modulation of conditions for ligand-receptor interaction

  • Original Articles
  • Published:
Inflammation Aims and scope Submit manuscript

Abstract

Results obtained with the luminol-dependent chemiluminescence (CL) technique show that with this technique, generation of oxygen radicals from an extra-as well as from an intracellular source is quantified. This investigation was performed in order to study the relationship between intra and extracellularly generated radicals in human granulocytes stimulated with the chemoattractant formyl-methionyl-leucyl-phenylalanine (FMLP). A difference in time course between extra and intracellular CL was observed. The extracellular response reached a maximum value after 1–2 min, whereas the intracellular response reached a maximum value after 5–7 min. The ED50 values for the two responses were the same, whereas the onset time was a little longer for the intracellular response. Both high and low concentrations of FMLP gave rise to CL. However, the ratio between the extra and the intracellular response differed depending on the concentration of FMLP; the ratio was decreased at low concentrations of FMLP. The same type of change was obtained when the ligand-receptor ratio was decreased through modulation of the number of exposed receptors. The ratio between extra and intracellular activities was also changed by cytochalasin B, removal of Ca2+, or removal of Na+. The role of the extra and intracellular oxygen radical production as well as possible regulatory mechanisms are discussed.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  1. Schiffman, E., B. A. Corcoran, andS. M. Whale. 1975. N-Formylmethionyl peptides as chemoattractants for leukocytes.Proc. Natl. Acad. Sci USA. 72:1059–1061.

    Google Scholar 

  2. Wilkinson, P. C. 1982. Chemotaxis and Inflammation. Churchill Livingstone, Edinburgh.

    Google Scholar 

  3. Klebanoff, S. J., andR. A. Clark. 1978. The Neutrophil. North Holland Publishing, Amsterdam.

    Google Scholar 

  4. Becker, E. L. 1979. A multifunctional receptor on the neutrophil for synthetic chemotactic oligopeptides.J. Reticuloendothel. Soc. 26:701–709.

    Google Scholar 

  5. Root, R. K., andM. S. Cohen. 1981. The micribicidal mechanisms of human neutrophils and eosinophils.Rev. Infect. Dis. 3:565–598.

    Google Scholar 

  6. Metcalf, J. A., J. I. Gallin, W. M. Nausseef, andR. K. Root. 1986. Laboratory Manual of Neutrophil Function. Raven Press, New York.

    Google Scholar 

  7. Movat, H. Z. 1985. The Inflammatory Reaction. Elsevier, Amsterdam.

    Google Scholar 

  8. Stendahl, O., B. I. Coble, C. Dahlgren, andJ. Hed. 1984. Myeloperoxidase modulates the phagocytic activity of polymorphonuclear neutrophil leukocytes. Studies with cells from a myeloperoxidase deficient patient.J. Clin. Invest. 73:366–373.

    Google Scholar 

  9. Clark, R. A., andS. Szot. 1982. Chemotactic factor inactivation by stimulated human neutrophils mediated by myeloperoxidase catalyzed methionine oxidation.J. Immunol. 128:1507–1513.

    Google Scholar 

  10. Bender, J. G., andD. E. VanEpps. 1982. Analysis of the bimodal chemiluminescence pattern stimulated in human neutrophils by chemotactic factors.Infect. Immun. 41:1062–1070.

    Google Scholar 

  11. Campbell, A. K., andM. B. Hallett. 1983. Measurement of intracellular calcium ions and oxygen radicals in polymorphonuclear leukocyte-erythrocyte ghost hybrids.J. Physiol. 338:537–550.

    Google Scholar 

  12. Dahlgren, C. 1987. Polymorphonuclear leukocyte chemiluminescence induced by formyl-methionyl-leucyl-phenylalanine and phorbol myristate acetate: Effects of catalase and super-oxide dismutase.Agents Actions 21:104–112.

    Google Scholar 

  13. Dahlgren, C. 1987. Difference in extracellular radical release after chemotactic factor and calcium ionophore activation of the oxygen radical generating system in human neutrophils.Biochim. Biophys. Acta 930:33–38.

    Google Scholar 

  14. Wymann, M. P., V. VonTscharner, D. A. Deranleau, andM. Baggiolini. 1986. A chemiluminescence assay for the real-time detection of HOOH released by stimulated neutrophils.J. Biolum. Chemilum. 1:144.

    Google Scholar 

  15. Boyum, A. 1968. Isolation of mononuclear cells and granulocytes from human blood. Isolation of mononuclear cells by one centrifugation and of granulocytes by combining centrifugation and sedimentation of 1×g.Scand. J. Clin. Lab. Invest. Suppl. 97:77–89.

    Google Scholar 

  16. Root, R. K., J. A. Metcalf, N. Oshino, andB. Chance. 1975. HOOH release from human granulocytes during phagocytosis I. Documentation, quantitation and some regulating factors.J. Clin. Invest. 55:945–955.

    Google Scholar 

  17. Dahlgren, C., T. Andersson, andO. Stendahl. 1987. Chemotactic factor binding and functional capacity: A comparison between human granulocytes and differentiated HL-60 cells.J. Leuk. Biol. 42:245–252.

    Google Scholar 

  18. Aniansson, H., O. Stendahl, andC. Dahlgren. 1984. Comparison between luminol and lucigenin dependent chemiluminescence of polymorphonuclear leukocytes.Acta Pathol. Microbiol. Scand. Sect. C. 92:357–361.

    Google Scholar 

  19. Dahlgren, C., andO. Stendahl. 1982. Effect of in vitro preincubation of polymorphonuclear leukocytes on formylmethionyl-leucyl-phenylalanine induced chemiluminescence.Infect. Immun. 37:34–39.

    Google Scholar 

  20. Dahlgren, C., andO. Stendahl. 1983. Role of myeloperoxidase in luminol dependent chemiluminescence of polymorphonuclear leukocytes.Infect. Immun. 39:736–741.

    Google Scholar 

  21. Dechatelet, L. R., G. D. Long, P. S. Shirley, D. A. Bass, M. J. Thomas, F. W. Henderson, andM. S. Cohen. 1982. Mechanism of the luminol-dependent chemiluminescence of human neutrophils.J. Immunol. 129:1589–1593.

    Google Scholar 

  22. Stevens, P., D. J. Winston, andK. VanDyke. 1978. In vitro evaluation of opsonic and cellular granulocyte function by luminol-dependent chemiluminescence: Utility in patients with severe neutropenia and cellular deficiency states.Infect. Immun. 22:41–51.

    Google Scholar 

  23. Ohno, Y., andJ. I. Gallin. 1985. Diffusion of extracellular hydrogen peroxide into intracellular compartments of human neutrophils.J. Biol. Chem. 260:8438–8446.

    Google Scholar 

  24. Root, R. K., andJ. A. Metcalf. 1977. H2O2 release from human granulocytes during phagocytosis.J. Clin. Invest. 60:1266–1279.

    Google Scholar 

  25. Dahlgren, C., andG. Briheim. 1985. Comparision between the luminol-dependent chemiluminescence of polymorphonuclear leukocytes and of the myeloperoxidase-HOOH system: Influence of pH, cations and protein.Photochem. Photobiol. 41:605–610.

    Google Scholar 

  26. Briheim, G., O. Stendahl, andC. Dahlgren. 1984. Intraand extracellular events in luminol-dependent chemiluminescence of polymorphonuclear leukocytes.Infect. Immun. 45:1–5.

    Google Scholar 

  27. Dahlgren, C., H. Aniansson, andK.-E. Magnusson. 1985. Pattern of formylmethionylleucyl-phenylalanine induced luminol- and lucigenin-dependent chemiluminescence in human neutrophils.Infect. Immun. 47:326–328.

    Google Scholar 

  28. McPhail, L. S., C. C. Clayton, andR. Snyderman. 1984. The NADPH oxidase of human polymorphonuclear leukocytes.J. Biol. Chem. 259:5768–5775.

    Google Scholar 

  29. Sklar, L. A. 1986. Ligand-receptor dynamics and signal amplification in the neutrophil.Adv. Immunol. 39:95–143.

    Google Scholar 

  30. Hallett, M. B., andA. K. Campbell. 1984. Is intracellular Ca++ the trigger for oxygen radical production by polymorphonuclear leucocytes?Cell Calcium 5:1–19.

    Google Scholar 

  31. Sklar, L. A., andZ. G. Oades. 1985. Signal transduction and ligand-receptor dynamics in the neutrophil.J. Biol. Chem. 260:11468–11475.

    Google Scholar 

  32. Lew, P. D., C. B. Wollheim, F. A. Waldvogel, andT. Pozzan, 1984. Modulation of cytosolic free calcium transients by changes in intracellular calcium buffering capacity: Correlation with exocytosis and O 2 production in human neutrophils.J. Cell Biol. 99:1212–1220.

    Google Scholar 

  33. Rossi, F., M. Grezeskowiak, andV. Dellabianca. 1986. Double stimulation with FMLP and Con A restores the activation of the respiratory burst but not the phosphinositide turnover in Ca++-depleted human neutrophils. A further example of dissociation between stimulation of the NADPH oxidase and phosphinositide turnover.Biochem. Biophys, Res. Commun. 140:1–11.

    Google Scholar 

  34. Pozzan, T., D. P. Lew, C. B. Wollheim, andR. Y. Tsien. 1983. Is cytosolic ionized calcium regulating neutrophil activation?Science 221:1413–1415.

    Google Scholar 

  35. Dahlgren, C., K.-E. Magnusson, O. Stendahl, andSundqvist, T. 1982. Modulation of polymorphonuclear leukocyte chemiluminescent response to the chemoattractant f-Met-Leu-Phe.Int. Arch. Allergy Appt. Immun. 68:79–83.

    Google Scholar 

  36. Fearon, D. T., andL. A. Collins. 1983. Increased expression of C3b receptors on poly-morphonuclear leukocytes induced by chemotactic factors and by purification procedures.J. Immunol. 130:370–375.

    Google Scholar 

  37. Andersson, T., C. Dahlgren, D. P. Lew, andO. Stendahl. 1987. Cell surface expression of f-Met-Leu-Phe receptors on human neutrophils.J. Clin. Invest. 79:1226–1233.

    Google Scholar 

  38. Zigmond, S. H., A. Woodworth, andG. Daukas. 1985. Effects of sodium on chemotactic peptide binding to polymorphonuclear leukocytes.J. Immunol. 135:531–536.

    Google Scholar 

  39. Borregaard, N., andA. Tauber. 1984. Subcellular localization of the human neutrophil NADPH oxidase:b-cytochrome and associated flavoprotein.J. Biol. Chem. 259:47–52.

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Dahlgren, C. Effects on extra- and intracellularly localized, chemoattractant-induced, oxygen radical production in neutrophils following modulation of conditions for ligand-receptor interaction. Inflammation 12, 335–349 (1988). https://doi.org/10.1007/BF00915769

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00915769

Keywords

Navigation