Skip to main content
SpringerLink
Log in

Changes in respiratory burst activity during human monocyte differentiation in suspension culture

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

Abstract

Monocytes undergo a process of differentiation following their accumulation into extravascular spaces. This process has been examined previously by culturing monocytes and identifying changes in cell morphology, metabolism, and function over time. The present study was designed to characterize mononuclear phagocyte respiratory burst activity as related to differentiation by measuring chemiluminescence and Superoxide anion generation in cultured human monocytes. Monocytes maintained in Teflon vials for up to 12 days increased in size, were positive for nonspecific esterase, and retained the ability to ingest latex particles. During culture, however, cells progressively lost their peroxidase-positive granules. When monocytes were cultured for one or five days, they elicited less than 50% of the luminol-enhanced chemiluminescence produced by fresh monocytes following PMA stimulation. By day 7, less than 20% of day 0 PMA-elicited chemiluminescence was observed. A comparable loss of serum-opsonized zymosan-induced chemiluminescence occurred during monocyte culture. Since it is recognized that luminol-enhanced chemiluminescence is, in large part, dependent upon myeloperoxidase and since differentiated mononuclear phagocytes are only minimally peroxidase-positive, cultured monocyte respiratory burst activity was also assessed by directly quantifying Superoxide anion generation. When monocytes were cultured for three or five days, they elicited 38% more Superoxide anion than did fresh monocytes following PMA stimulation. At day 7, PMA-induced Superoxide anion release was comparable to day 0 levels. These data indicate that monocytes allowed to differentiate under nonadherent conditions maintain the ability to undergo a respiratory burst response as measured by Superoxide anion release, but they concomitantly lose peroxidase-dependent luminol-enhanced chemiluminescence. In this regard, monocytes cultured in suspension metabolically resemble macrophages that have undergone differentiation within sites of inflammation.

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

Access this article

Log in via an institution

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

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Van Furth R., J. W. M.van der Meer, A. B.van Oud Alblas, and W.Sluiter. 1982. Development of mononuclear phagocytes.In Self-Defense Mechanisms: Role of Macrophages. D. Mizuno, Z. A. Cohn, K. Takeya, and N. Ishida, editors. 25–43.

  2. van der Rhee, H. J., C. P. M. van der Burgh-deWinter, andW. Th. Daems. 1979. The differentiation of monocytes into macrophages, epithelioid cells, and multinucleated giant cells in subcutaneous granulomas. I. Fine structure.Cell Tissue Res. 197:355–378.

    Google Scholar 

  3. Oren, R., A. E. Farnham, K. Saito, E. Milofsky, andM. L. Karnovsky. 1963. Metabolic patterns in three types of phagocytizing cells.J. Cell Biol. 17:487–501.

    Google Scholar 

  4. Davies, P., andR. J. Bonney. 1979. Secretory products of mononuclear phagocytes: A brief review.J. Reticuloendothel. Soc. 26:37–47.

    Google Scholar 

  5. Rinehart, J. J., R. Vessella, P. Lange, M. E. Kaplan, andB. J. Gormus. 1979. Characterization and comparison of human monocyte- and macrophage-induced tumor cell cytotoxicity.J. Lab. Clin. Med. 96:361–369.

    Google Scholar 

  6. Biondi, A., T. H. Rossing, J. Bennett, andR. F. Todd, III. 1984. Surface membrane heterogeneity among human mononuclear phagocytes.J. Immunol. 132:1237–1243.

    Google Scholar 

  7. Johnston, R. B., Jr., C. A. Godzik, andZ. A. Cohn. 1978. Increased superoxide anion production by immunologically activated and chemically elicited macrophages.J. Exp. Med. 148:115–127.

    Google Scholar 

  8. Alpert, S. E., H. S. Auerbach, F. S. Cole, andH. R. Colten. 1983. Macrophage maturation: Differences in complement secretion by marrow, monocyte, and tissue macrophages detected with an improved hemolytic plaque assay.J. Immunol. 130:102–107.

    Google Scholar 

  9. Meuret, G., O. Schildknecht, P. Joder, andH. Senn. 1980. Proliferative activity and bacteriostatic potential of human blood monocytes, macrophages in pleural effusions, ascites, and of alveolar macrophages.Blut 40:17–25.

    Google Scholar 

  10. van Furth, R., J. A. Raeburn, andT. L. van Zwet, 1979. Characteristics of human mononuclear phagocytes.Blood 54:488–500.

    Google Scholar 

  11. Bennett, W. E., andZ. A. Cohn. The isolation and selected properties of blood monocytes. 1966.J. Exp. Med. 123:145–159.

    Google Scholar 

  12. Johnson, W. D., B. Mei, andZ. A. Cohn. 1972. The separation, long-term cultivation, and maturation of the human monocyte.J. Exp. Med. 146:1613–1626.

    Google Scholar 

  13. Zuckerman, S. H., S. K. Ackerman, andS. D. Douglas. 1979. Long-term human peripheral blood monocyte cultures: Establishment, metabolism, and morphology of primary human monocyte-macrophage cell cultures.Immunology 38:401–411.

    Google Scholar 

  14. Andreesen, R., J. Picht, andG. W. Löhr. 1983. Primary cultures of human blood-borne macrophages grown on hydrophobic Teflon membranes.J. Immunol. Methods 56:295–304.

    Google Scholar 

  15. van der Meer, J. W. M., J. S. van de Gevel, I. Elzenga-Claassen, andR. van Furth. 1979. Suspension cultures of mononuclear phagocytes in the Teflon culture bag.Cell. Immunol. 42:208–212.

    Google Scholar 

  16. van der Meer, W. M., J. S. van de Gevel, A. B. van Oud Alblas, J. A. Kramps, L. van Zwet, P. C. J. Leijh, andR. van Furth. 1982. Characteristics of human monocytes cultured in the Teflon culture bag.Immunology 47:617–625.

    Google Scholar 

  17. Zeller, J. M., A. L. Landay, T. F. Lint, andH. Gewurz. 1986. Enhancement of human peripheral blood monocyte respiratory burst activity by aggregated C-reactive protein.J. Leukocyte Biol. 40:769–783.

    Google Scholar 

  18. Wright, S. D., andS. C. Silverstein. 1982. Tumor-promoting phorbol esters stimulate C3b and C3b' receptor-mediated phagocytosis in cultured human monocytes.J. Exp. Med. 156:1149–1164.

    Google Scholar 

  19. Ross, G. D., andR. J. Winchester. 1980. Methods for enumerating lymphocyte populations.In Manual of Clinical Immunology. N. R. Rose and H. Friedman, editors. American Society of Microbiology, Washington, D.C. 213.

    Google Scholar 

  20. Miller, G. A., andP. S. Morahan. 1981. Use of non-specific esterase stain.In Methods for Studying Mononuclear Phagocytes. D. O. Adams, P. J. Edelson, and H. S. Koren, editors. Academic Press, New York. 367–374.

    Google Scholar 

  21. Meltzer, M.S. 1981. Use of peroxidase stain by the Kaplow method.In Methods for Studying Mononuclear Phagocytes. D. O. Adams, P. J. Edelson, and H. S. Koren, editors. Academic Press, New York. 363–366.

    Google Scholar 

  22. Pick, E., andD. Mizel. 1981. Rapid microassays for the measurement of superoxide and hydrogen peroxide production by macrophages in culture using an automatic enzyme immunoassay reader.J. Immunol. Methods 46:211–226.

    Google Scholar 

  23. Musson, R. A., L. C. McPhail, H. Shafran, andR. B. Johnston, Jr. 1982. Differences in the ability of human peripheral blood monocytes and in vitro monocyte derived macrophages to produce Superoxide anion: Studies with cells from normals and patients with chronic granulomatous disease.J. Reticuloendothel. Soc. 31:261–266.

    Google Scholar 

  24. Nakagawara, A., C. F. Nathan, andZ. A. Cohn. 1981. Hydrogen peroxide metabolism in human monocytes during differentiation in vitro.J. Clin. Invest. 68:1243–1252.

    Google Scholar 

  25. Turpin, J., E. M. Hersh, andG. Lopez-Berestein. 1986. Characterization of small and large human peripheral blood monocytes: Effects of in vitro maturation on hydrogen peroxide release and on the response to macrophage activators.J. Immunol. 136:4194–4198.

    Google Scholar 

  26. Johnston, R. B., Jr., J. E. Lehmeyer, andL. A. Guthrie. 1976. Generation of Superoxide anion and chemiluminescence by human monocytes during phagocytosis and on contact with surface-bound immunoglobulin G.J. Exp. Med. 143:1551–1556.

    Google Scholar 

  27. Kitagawa, S., F. Takaku, andS. Sakamoto. 1980. Evidence that proteases are involved in superoxide production by human polymorphonuclear leukocytes and monocytes.J. Clin. Invest. 65:74–81.

    Google Scholar 

  28. Biggar, W. D., S. Buron, andB. Holmes. 1976. Bactericidal mechanisms in rabbit alveolar macrophages: Evidence against peroxidase and hydrogen peroxide bactericidal mechanisms.Infect. Immun. 14:6–10.

    Google Scholar 

  29. DeChatelet, L. R., D. Mullikin, andC. E. McCall. 1975. The generation of superoxide anion by various types of phagocyte.J. Infect. Dis. 131:443–446.

    Google Scholar 

  30. Nathan, C. F., andR. K. Root. 1977. Hydrogen peroxide release from mouse peritoneal macrophages. Dependence on sequential activation and triggering.J. Exp. Med. 146:1648–1662.

    Google Scholar 

  31. Seim, S. 1982. Production of reactive oxygen species and chemiluminescence by human monocytes during differentiation and lymphokine activation in vitro.Acta Pathol. Microbiol. Immunol. Scand. Sect. C 90:179–185.

    Google Scholar 

  32. Hammerstrom, J. 1979. Human macrophage differentiation in vivo and in vitro.Acta Pathol. Microbiol. Scand. Sect. C 87:113–120.

    Google Scholar 

  33. Kaplan, G., andG. Gaudernack. 1982. In vitro differentiation of human monocytes. Differences in monocyte phenotypes induced by cultivation on glass or on collagen.J. Exp. Med. 156:1101–1114.

    Google Scholar 

  34. Seim, S. 1983. Role of myeloperoxidease in the luminol-dependent chemiluminescence response of phagocytosing human monocytes.Acta Pathol. Microbiol. Immunol. Scand. Sect. C 91:123–128.

    Google Scholar 

  35. Allen, R. C. 1986. Phagocytic leukocyte oxygenation activities and chemiluminescence: A kinetic approach to analysis.Methods Enzymol. 133:449–493.

    Google Scholar 

  36. Rosen, H., andS. J. Klebanoff. 1976. Chemiluminescence and Superoxide production by myeloperoxidase-deficient leukocytes.J. Clin. Invest. 58:50–60.

    Google Scholar 

  37. Berton, G., andS. Gordon. 1983. Superoxide release by peritoneal and bone marrow-derived mouse macrophages. Modulation by adherence and cell activation.Immunology 49:693–704.

    Google Scholar 

  38. Pabst, M. J., H. B. Hedegaard, andR. B. Johnston, Jr. 1982. Cultured monocytes require exposure to bacterial products to maintain an optimal oxygen radical response.J. Immunol. 128:123–128.

    Google Scholar 

  39. Cohen, M. S., D. E. Mesler, R. G. Snipes, andT. K. Gray. 1986. 1,25-Dihydroxyvitamin D3 activates secretion of hydrogen peroxide by human monocytes.J. Immunol. 136:1049–1053.

    Google Scholar 

  40. Nakagawara, A., N. M. Desantis, N. Nogueira, andC. F. Nathan. 1982. Lymphokines enhance the capacity of human monocytes to secrete reactive oxygen intermediates.J. Clin. Invest. 70:1042–1048.

    Google Scholar 

  41. Cassatella, M. A., V. Della Bianca, G. Berton, andF. Rossi. 1985. Activation by gamma Interferon of human macrophage capability to produce toxic oxygen molecules is accompanied by a decreasedK m of the superoxide-generating NADPH oxidase.Biochim. Biophys. Acta 132:908–914.

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Medical Nursing, Rush-Presbyterian-St. Luke's Medical Center, 60612, Chicago, Illinois

    Janice M. Zeller

  2. Department of Immunology/Microbiology, Rush-Presbyterian-St. Luke's Medical Center, 60612, Chicago, Illinois

    Janice M. Zeller, Janice Caliendo & Thomas F. Lint

  3. Department of Medicine, University of Chicago, 60637, Chicago, Illinois

    Deborah J. Nelson

Authors
  1. Janice M. Zeller
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Janice Caliendo
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Thomas F. Lint
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Deborah J. Nelson
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

Presented, in part, to the Reticuloendothelial Society, August 1985.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Zeller, J.M., Caliendo, J., Lint, T.F. et al. Changes in respiratory burst activity during human monocyte differentiation in suspension culture. Inflammation 12, 585–595 (1988). https://doi.org/10.1007/BF00914320

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00914320

Keywords

Search

Navigation