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.
Similar content being viewed by others
References
Schiffman, E., B. A. Corcoran, andS. M. Whale. 1975. N-Formylmethionyl peptides as chemoattractants for leukocytes.Proc. Natl. Acad. Sci USA. 72:1059–1061.
Wilkinson, P. C. 1982. Chemotaxis and Inflammation. Churchill Livingstone, Edinburgh.
Klebanoff, S. J., andR. A. Clark. 1978. The Neutrophil. North Holland Publishing, Amsterdam.
Becker, E. L. 1979. A multifunctional receptor on the neutrophil for synthetic chemotactic oligopeptides.J. Reticuloendothel. Soc. 26:701–709.
Root, R. K., andM. S. Cohen. 1981. The micribicidal mechanisms of human neutrophils and eosinophils.Rev. Infect. Dis. 3:565–598.
Metcalf, J. A., J. I. Gallin, W. M. Nausseef, andR. K. Root. 1986. Laboratory Manual of Neutrophil Function. Raven Press, New York.
Movat, H. Z. 1985. The Inflammatory Reaction. Elsevier, Amsterdam.
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.
Clark, R. A., andS. Szot. 1982. Chemotactic factor inactivation by stimulated human neutrophils mediated by myeloperoxidase catalyzed methionine oxidation.J. Immunol. 128:1507–1513.
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.
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.
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.
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.
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.
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.
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.
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.
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.
Dahlgren, C., andO. Stendahl. 1982. Effect of in vitro preincubation of polymorphonuclear leukocytes on formylmethionyl-leucyl-phenylalanine induced chemiluminescence.Infect. Immun. 37:34–39.
Dahlgren, C., andO. Stendahl. 1983. Role of myeloperoxidase in luminol dependent chemiluminescence of polymorphonuclear leukocytes.Infect. Immun. 39:736–741.
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.
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.
Ohno, Y., andJ. I. Gallin. 1985. Diffusion of extracellular hydrogen peroxide into intracellular compartments of human neutrophils.J. Biol. Chem. 260:8438–8446.
Root, R. K., andJ. A. Metcalf. 1977. H2O2 release from human granulocytes during phagocytosis.J. Clin. Invest. 60:1266–1279.
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.
Briheim, G., O. Stendahl, andC. Dahlgren. 1984. Intraand extracellular events in luminol-dependent chemiluminescence of polymorphonuclear leukocytes.Infect. Immun. 45:1–5.
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.
McPhail, L. S., C. C. Clayton, andR. Snyderman. 1984. The NADPH oxidase of human polymorphonuclear leukocytes.J. Biol. Chem. 259:5768–5775.
Sklar, L. A. 1986. Ligand-receptor dynamics and signal amplification in the neutrophil.Adv. Immunol. 39:95–143.
Hallett, M. B., andA. K. Campbell. 1984. Is intracellular Ca++ the trigger for oxygen radical production by polymorphonuclear leucocytes?Cell Calcium 5:1–19.
Sklar, L. A., andZ. G. Oades. 1985. Signal transduction and ligand-receptor dynamics in the neutrophil.J. Biol. Chem. 260:11468–11475.
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.
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.
Pozzan, T., D. P. Lew, C. B. Wollheim, andR. Y. Tsien. 1983. Is cytosolic ionized calcium regulating neutrophil activation?Science 221:1413–1415.
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.
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.
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.
Zigmond, S. H., A. Woodworth, andG. Daukas. 1985. Effects of sodium on chemotactic peptide binding to polymorphonuclear leukocytes.J. Immunol. 135:531–536.
Borregaard, N., andA. Tauber. 1984. Subcellular localization of the human neutrophil NADPH oxidase:b-cytochrome and associated flavoprotein.J. Biol. Chem. 259:47–52.
Author information
Authors and Affiliations
Rights 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
Issue Date:
DOI: https://doi.org/10.1007/BF00915769