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Ultrastructural localization of NADPH-oxidase activity in murine peritoneal macrophages during phagocytosis ofBrucella

Correlation with the production of Superoxide anions

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Virchows Archiv B

Summary

The localization of the enzyme NADPH oxidase in mouse peritoneal macrophages unstimulated or after phagocytosis ofBruceila suis was investigated by electron microscopy in normal mice and mice immunized againstB. suis. The enzyme was clearly visualized on mitochondrial cristae, smooth endoplasmic reticulum, and the plasma membrane; its activity correlated mainly with the state of the endoplasmic reticulum which itself reflected macrophage activation. The enzyme turn-over appeared to be accelerated after phagocytosis; the phagosome mambrane was seldom stained by the enzyme reaction.

These macrophages were also examined for the production of super-oxide anions in vitro, either unstimulated or after phagocytosis. Phagocytosis increased the production of Superoxide anions, especially in immunized animals. These results are discussed with regard to the role that the products of oxidative metabolism play in the inactivation of bacteria by phagocytic cells.

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References

  • Ago, Y, Ogawa K (1971) Ultrastructural demonstration of NADPH ferricyanide reductase activity. Acta Histochem 4:161–165

    CAS  Google Scholar 

  • Babior BM (1978) Oxygen-dependent killing by phagocytes. N Engl J Med 298:659–668

    PubMed  CAS  Google Scholar 

  • Babior B, Kipnes R, Curnutte J (1973) Biological defense mechanisms. The production by leukocytes of Superoxide, a potent bactericidal agent. J Clin Invest 52:741–744

    Article  PubMed  CAS  Google Scholar 

  • Chatelet LR de (1978) Initiation of respiratory burst in human polymorphonuclear neutrophils; a critical review. J Reticuloendothel Soc 24:73–91

    Google Scholar 

  • Dreath DD, Karnovsky ML (1975) Superoxide production by phagocytic leucocytes. J Exp Med 141:257–262

    Article  Google Scholar 

  • Fitzgeorge RB, Keppie J, Smitz H (1965) The relation between resistance to hydrogen peroxide and virulence inBrucellae. J Pathol 89:745–747

    Article  CAS  Google Scholar 

  • Gay B, Mauss H, Sanchez-Teff S (1981) Aspects ultrastructuraux de la phagocytosein vivo etin vitro deBrucella par les macrophages du péritoine de la souris. Ann Immunol [Paris] 132D:299–313

    Google Scholar 

  • Goldstein IM, Cerqueira M, Lind S, Kaplan H (1977) Evidence that the Superoxide generating system of human granulocytes is associated with the cell surface. J Clin Invest 59:249–254

    Article  PubMed  CAS  Google Scholar 

  • Hanker JJ (1975) Oxidoreductases. In: M. Hayat (ed) Electron microscopy of enzymes. Principles and methods. Van Nostrand Reinhold, New York, p 69

    Google Scholar 

  • Hsiu-San Lin, Gordon S (1979) Secretion of plasminogen activator by bone marrow-derived mononuclear phagocytes and its enhancement by colony stimulating factor. J Exp Med 150:231–245

    Article  Google Scholar 

  • Iverson DB, Iverson PW, Spitznagel JK, Chatelet LR de (1978) Subcellular localization of NAD(P)H-oxidase(s) in human neutrophilic polymorphonuclear leucocytes. Biochem J 176:175–178

    PubMed  CAS  Google Scholar 

  • Johnson RB, Godzyk CA, Cohn ZA (1978) Increased Superoxide anion production by immunologically activated and chemically elicited macrophages. J Exp Med 148:115–127

    Article  Google Scholar 

  • Kerpel-Fronius S, Hajos F (1970) An attempt to evaluate the specificity of the fine structural localization of oxidoreductase activity with a simultaneous coupling method. J Histochem Cytochem 18:219–220

    PubMed  CAS  Google Scholar 

  • Lopez-Merino A, Asselineau A, Serre A, Roux J, Bascoul S, Lacave C (1976) Immunization by an insoluble fraction extracted fromBrucella melitensis. Immunological and chemical characterization of the active substances. Infect Immun 13:311–321

    PubMed  CAS  Google Scholar 

  • Luft JH (1961) Improvement in epoxy-embedding methods. J Biophys Biochem Cytol 9:409–414

    Article  PubMed  CAS  Google Scholar 

  • Murray HW, Cohn ZA (1980) Macrophage oxygen-dependant antimicrobial activity. III. Enhanced oxidative metabolism as an expression of macrophage activation. J Exp Med 152:1516–1609

    Google Scholar 

  • Patriarca P, Kakinuma K, Tedesco P, Rossi F (1975) Studies on the mechanism of metabolic stimulation of polymorphonuclear leucocytes during phagocytosis. I. Evidence for superoxide anion involvement in the production of NADPH. Biochim Biophys Acta 385:380–386

    PubMed  CAS  Google Scholar 

  • Romeo D, Zabucchi G, Marzi T, Rossi F (1975) Kinetic and enzymatic features of metabolic stimulation of alveolar and peritoneal macrophages challenged with bacteria. Exp Cell Res 78:423–432

    Article  Google Scholar 

  • Root RK, Cohen MS (1981) The microbicidal mechanisms of human neutrophils and eosinophils. Rev Infect Dis 3:565–598

    PubMed  CAS  Google Scholar 

  • Rossi F, Zatti M (1964) Biochemical aspects of phagocytosis in polynuclear phagocytes. NAD and NADPH oxidation by the granules of resting and phagocytizing cells. Experientia 20:21–23

    Article  PubMed  CAS  Google Scholar 

  • Roux J, Asselineau J, Serre A, Lacave C (1967) Propriétés immunologiques d’un extrait phenolinsoluble deB. melitensis (fraction P.I.). Ann Immunol [Paris] 113:411–423

    CAS  Google Scholar 

  • Stach JL, Fumoux F, Strobel M, Baret A, Michelson M (1981) Augmentation de l’activité dismutasique dans les leucocytes spléniques de souris sensibles au B.C.G. et àMycobacterium lepraemurium. CR Acad Sci Paris [Ser. III] 293:573–578

    Google Scholar 

  • Takanaka K, O’Brien PJ (1975) Mechanism of H2O2 Production by leucocytes. Properties of the NAD(P)H-oxidase activity of intact neutrophils. Arch Biochem Biophys 169:428–435

    Article  PubMed  CAS  Google Scholar 

  • Tauber AL, Babior BM (1977) Evidence for hydroxyl radical production by human neutrophils. J Clin Invest 60:374–379

    Article  PubMed  CAS  Google Scholar 

  • Weiss SJ, King GW, Lo Buglio AF (1977) Evidence for hydroxyl radical generation by human monocytes. J Clin Invest 60:370–374

    Article  PubMed  CAS  Google Scholar 

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Gay, B., Sanchez-Teff, S. & Caravano, R. Ultrastructural localization of NADPH-oxidase activity in murine peritoneal macrophages during phagocytosis ofBrucella . Virchows Archiv B Cell Pathol 45, 147–155 (1984). https://doi.org/10.1007/BF02889861

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  • DOI: https://doi.org/10.1007/BF02889861

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