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. 1978 Jul 1;148(1):115–129. doi: 10.1084/jem.148.1.115

Increased superoxide anion production by immunologically activated and chemically elicited macrophages

Jr RB Johnston, CA Godzik, ZA Cohn
PMCID: PMC2184904  PMID: 209122

Abstract

We studied the capacity of cultured mouse peritoneal macrophages to generate superoxide anion (O(2-)), the initial product of conversion of oxygen to microbicidal species, during phagocytosis of opsonized zymosan or upon contact with the membrane-active agent phorbel myristate acetate (PMA). Macrophages from mice infected with Bacille Calmette-Guerin (BCG) or injected intraperitoneally with thioglycollate broth or endotoxin, released up to 12 times more O(2-) than did resident peritoneal macrophages, depending upon the cell type and whether the stimulus was zymosan or PMA. There was little if any O(2-) release from resting (unstimulated) macrophages. The density of cells on culture dishes was an important variable since crowding of the dish markedly reduced the efficiency of O(2-) production. The enhanced O(2-) release of chemically elicited and infection-activated macrophages was noted after stimulation with a wide range of concentrations of PMA and zymosan, at all time points studied (up to 120 min), and with cells maintained for 140 rain to 16 days in culture. The O(2-) response of resident cells improved twofold to zymosan and ninefold to PMA during the first 3 days in culture. The capacity to release O~ appears to be limited to actively phagocytic cell types: murine macrophage-like tumor lines and cultured human monocytes released O(2-) when stimulated by PMA or zymosan, fibroblast and endothelial lines and embryo-derived cells did not. Activity of superoxide dismutase, which removes O(2-), was not detectable in culture supernates of any cell type, and thus, differences in detectable O(2-) could not be attributed to variations in the release of this enzyme. We conclude that the phagocytosis- associated respiratory burst is significantly enhanced in mononuclear phagocytes obtained ai~r chemical inflammation or BCG infection. Increased capacity to generate O(2-) and other oxygen radicals during phagocytosis could contribute to the improved microbicidal and tumoricidal activity of activated macrophages.

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Selected References

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  1. Babior B. M., Curnutte J. T., McMurrich B. J. The particulate superoxide-forming system from human neutrophils. Properties of the system and further evidence supporting its participation in the respiratory burst. J Clin Invest. 1976 Oct;58(4):989–996. doi: 10.1172/JCI108553. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Baehner R. L., Johnston R. B., Jr Monocyte function in children with neutropenia and chronic infections. Blood. 1972 Jul;40(1):31–41. [PubMed] [Google Scholar]
  3. Blanden R. V. Modification of macrophage function. J Reticuloendothel Soc. 1968 Jun;5(3):179–202. [PubMed] [Google Scholar]
  4. COHN Z. A., BENSON B. THE DIFFERENTIATION OF MONONUCLEAR PHAGOCYTES. MORPHOLOGY, CYTOCHEMISTRY, AND BIOCHEMISTRY. J Exp Med. 1965 Jan 1;121:153–170. doi: 10.1084/jem.121.1.153. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. DeChatelet L. R., McPhail L. C., Mullikin D., McCall C. E. An isotopic assay for NADPH oxidase activity and some characteristics of the enzyme from human polymorphonuclear leukocytes. J Clin Invest. 1975 Apr;55(4):714–721. doi: 10.1172/JCI107981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Drath D. B., Karnovsky M. L. Superoxide production by phagocytic leukocytes. J Exp Med. 1975 Jan 1;141(1):257–262. doi: 10.1084/jem.141.1.257. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Evans D. G., Myrvik Q. N. Biology of the mycobacterioses. Studies on glucose oxidation during the interaction of alveolar macrophages and bacteria. Ann N Y Acad Sci. 1968 Sep 5;154(1):167–176. doi: 10.1111/j.1749-6632.1968.tb16707.x. [DOI] [PubMed] [Google Scholar]
  8. Evans R. Macrophage-mediated cytotoxicity: its possible role in rheumatoid arthritis. Ann N Y Acad Sci. 1975 Jun 13;256:275–287. doi: 10.1111/j.1749-6632.1975.tb36054.x. [DOI] [PubMed] [Google Scholar]
  9. Gordon S. Macrophage neutral proteinases and chronic inflammation. Ann N Y Acad Sci. 1976;278:176–189. doi: 10.1111/j.1749-6632.1976.tb47028.x. [DOI] [PubMed] [Google Scholar]
  10. Hohn D. C., Lehrer R. I. NADPH oxidase deficiency in X-linked chronic granulomatous disease. J Clin Invest. 1975 Apr;55(4):707–713. doi: 10.1172/JCI107980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Johnson W. D., Jr, Mei B., Cohn Z. A. The separation, long-term cultivation, and maturation of the human monocyte. J Exp Med. 1977 Dec 1;146(6):1613–1626. doi: 10.1084/jem.146.6.1613. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Johnston R. B., Jr, Keele B. B., Jr, Misra H. P., Lehmeyer J. E., Webb L. S., Baehner R. L., RaJagopalan K. V. The role of superoxide anion generation in phagocytic bactericidal activity. Studies with normal and chronic granulomatous disease leukocytes. J Clin Invest. 1975 Jun;55(6):1357–1372. doi: 10.1172/JCI108055. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Johnston R. B., Jr, Lehmeyer J. E., Guthrie L. A. Generation of superoxide anion and chemiluminescence by human monocytes during phagocytosis and on contact with surface-bound immunoglobulin G. J Exp Med. 1976 Jun 1;143(6):1551–1556. doi: 10.1084/jem.143.6.1551. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Karnovsky M. L., Lazdins J., Drath D., Harper A. Biochemical characteristics of activated macrophages. Ann N Y Acad Sci. 1975 Jun 13;256:266–274. doi: 10.1111/j.1749-6632.1975.tb36053.x. [DOI] [PubMed] [Google Scholar]
  15. Keller R. Mechanisms by which activated normal macrophages destroy syngeneic rat tumour cells in vitro. Cytokinetics, non-involvement of T lymphocytes, and effect of metabolic inhibitors. Immunology. 1974 Aug;27(2):285–298. [PMC free article] [PubMed] [Google Scholar]
  16. Klebanoff S. J. Antimicrobial mechanisms in neutrophilic polymorphonuclear leukocytes. Semin Hematol. 1975 Apr;12(2):117–142. [PubMed] [Google Scholar]
  17. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  18. Leake E. S., Myrvik Q. N. Changes in morphology and in lysozyme content of free alveolar cells after the intravenous injection of killed BCG in oil. J Reticuloendothel Soc. 1968 Feb;5(1):33–53. [PubMed] [Google Scholar]
  19. McCord J. M., Fridovich I. Superoxide dismutase. An enzymic function for erythrocuprein (hemocuprein). J Biol Chem. 1969 Nov 25;244(22):6049–6055. [PubMed] [Google Scholar]
  20. McPhail L. C., DeChatelet L. R., Shirley P. S. Further characterization of NADPH oxidase activity of human polymorphonuclear leukocytes. J Clin Invest. 1976 Oct;58(4):774–780. doi: 10.1172/JCI108528. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Nathan C. F., Karnovsky M. L., David J. R. Alterations of macrophage functions by mediators from lymphocytes. J Exp Med. 1971 Jun 1;133(6):1356–1376. doi: 10.1084/jem.133.6.1356. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Nathan C. F., Root R. K. Hydrogen peroxide release from mouse peritoneal macrophages: dependence on sequential activation and triggering. J Exp Med. 1977 Dec 1;146(6):1648–1662. doi: 10.1084/jem.146.6.1648. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Nelson R. D., Mills E. L., Simmons R. L., Quie P. G. Chemiluminescence response of phagocytizing human monocytes. Infect Immun. 1976 Jul;14(1):129–134. doi: 10.1128/iai.14.1.129-134.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Patriarca P., Cramer R., Moncalvo S., Rossi F., Romeo D. Enzymatic basis of metabolic stimulation in leucocytes during phagocytosis: the role of activated NADPH oxidase. Arch Biochem Biophys. 1971 Jul;145(1):255–262. doi: 10.1016/0003-9861(71)90034-8. [DOI] [PubMed] [Google Scholar]
  25. Ratzan K. R., Musher D. M., Keusch G. T., Weinstein L. Correlation of increased metabolic activity, resistance to infection, enhanced phagocytosis, and inhibition of bacterial growth by macrophages from Listeria- and BCG-infected mice. Infect Immun. 1972 Apr;5(4):499–504. doi: 10.1128/iai.5.4.499-504.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Romeo D., Zabucchi G., Marzi T., Rossi F. Kinetic and enzymatic features of metabolic stimulation of alveolar and peritoneal macrophages challenged with bacteria. Exp Cell Res. 1973 Apr;78(2):423–432. doi: 10.1016/0014-4827(73)90087-6. [DOI] [PubMed] [Google Scholar]
  27. Sagone A. L., Jr, King G. W., Metz E. N. A comparison of the metabolic response to phagocytosis in human granulocytes and monocytes. J Clin Invest. 1976 May;57(5):1352–1358. doi: 10.1172/JCI108403. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Stubbs M., Kühner A. V., Glass E. A., David J. R., Karnovsky M. L. Metabolic and functonal studies on activated mouse macrophages. J Exp Med. 1973 Feb 1;137(2):537–542. doi: 10.1084/jem.137.2.537. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Weiss S. J., King G. W., LoBuglio A. F. Evidence for hydroxyl radical generation by human Monocytes. J Clin Invest. 1977 Aug;60(2):370–373. doi: 10.1172/JCI108785. [DOI] [PMC free article] [PubMed] [Google Scholar]

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