, Volume 4, Issue 3, pp 209–221 | Cite as

Endotoxin induces the expression of prostaglandin H synthase-2 and eicosanoid formation in cells of the human monocytic cell line Mono Mac 6

  • T. Christoph
  • M. Widerna
  • A. Bodenteich
  • J. Berg


In controlling inflammation, monocytes and macrophages not only secrete various cytokines but also eicosanoids. Since the human monocytic cell line, Mono Mac 6, represents cells at the stage of mature blood monocytes, cells of this line were investigated to find out whether they also resemble mature monocytes with respect to inflammation. In response to lipopolysaccharide (LPS), Mono Mac 6 cells secreted large amounts of eicosanoids, 6-keto-prostaglandin F (6k-PGF), thromboxane B2 (TXB2) and prostaglandin E2 (PGE2). The considerable increase in eicosanoid secretion is attributed to the expression of the inducible isoform of the prostaglandin H synthase (PGHS-2) since the PGHS-2-specific steady-state mRNA level, PGHS-2 protein, and the enzyme activity of PGHS were strikingly increased in the absence of detectable PGHS-1 protein. PGHS-2 expression was inhibited by either dexamethasone or cycloheximide. The non-steroidal anti-inflammatory drug (NSAID) diclofenac inhibited the increased PGHS activity. The expression of the PGHS-2 gene was accompanied by the formation of TNFα, IL-6 and IL-8, but not by IL-1β.


monocytes differentiation PGHS-2 cytokines 


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  1. 1.
    Beelen RHJ, Betjes MGH, Kamperdijk EWA. Macrophages: general aspects. In: Bruijnzeel-Koomen CAFM, Hoefsmit ECM, eds. Immunopharmacology of Macrophages and other Antigen-presenting Cells. London: Academic Press; 1994;1–6.Google Scholar
  2. 2.
    Herschman HR. Regulation of prostaglandin synthase-1 and prostaglandin synthase-2. Cancer Metastasis Rev. 1994;13:241–56.PubMedCrossRefGoogle Scholar
  3. 3.
    Masferrer JL, Reddy ST, Zweifel BS et al. In vivo glucocorticoids regulate cyclooxygenase-2 but not cyclooxygenase-1 in peritoneal macrophages. J Pharmacol Exp Ther. 1994;270:1340–4.PubMedGoogle Scholar
  4. 4.
    Sano H, Hla T, Maier JAM et al. In vivo cyclooxygenase expression in synovial tissues of patients with rheumatoid arthritis and osteoarthritis in rats with adjuvant and streptococcal cell wall arthritis. J Clin Invest. 1992;89:97–108.PubMedGoogle Scholar
  5. 5.
    Seibert K, Zhang Y, Leahy K et al. Pharmacological and biochemical demonstration of the role of cyclooxygenase 2 in inflammation and pain. Proc Natl Acad Sci USA. 1994;91:12013–17.PubMedCrossRefGoogle Scholar
  6. 6.
    Vane JR, Mitchell JA, Appleton I et al. Inducible isoforms of cyclooxygenase and nitric-oxide synthase in inflammation. Proc Natl Acad Sci USA. 1994;91:2046–50.PubMedCrossRefGoogle Scholar
  7. 7.
    Patrignani P, Panara M, Greco A et al.. Biochemical and pharmacological characterisation of the cyclooxygenase activity of human blood prostaglandin endoperoxide synthases. J Pharmacol Exp Ther. 1994;271:1705–12.PubMedGoogle Scholar
  8. 8.
    Fu J-Y, Masferrer JL, Seibert K, Raz A, Needleman P. The induction and suppression of prostaglandin H2 synthase in human monocytes. J Biol Chem. 1990;265:16737–40.PubMedGoogle Scholar
  9. 9.
    Arias-Negrete S, Keller K, Chadee K. Proinflammatory cytokines regulate cyclooxygenase-2 mRNA expression in human macrophages. Biochem Biophys Res Commun. 1995;208:582–9.PubMedCrossRefGoogle Scholar
  10. 10.
    Hoff T, DeWitt DL, Kaever V, Resch K, Goppelt-Struebe M. Differentiation-associated expression of prostaglandin G/H synthase in monocytic cells. FEBS Lett. 1993;320:38–42.PubMedCrossRefGoogle Scholar
  11. 11.
    Koehler L, Hass R, DeWitt DL, Resch K, Goppelt-Struebe M. Glucocorticoid-induced reduction of prostanoid synthesis in TPA-differentiated U937 cells mainly due to reduced cyclooxygenase activity. Biochem Pharmacol. 1990;40:1307–16.PubMedCrossRefGoogle Scholar
  12. 12.
    Sanduja SK, Metha K, Xu X-M, Hsu S-M, Sanduja R, Wu KK. Differentiation-associated expression of prostaglandin H and thromboxane A synthases in monocytoid leukemia cell lines. Blood. 1991;78:3178–85.PubMedGoogle Scholar
  13. 13.
    Ziegler-Heitbrock HWL, Thiel E, Fütterer A, Herzog V, Wirtz A, Wirtz A, Riethmüller G. Establishment of a human cell line (MONO MAC 6) with characteristics of mature monocytes. Int J Cancer. 1988;41:456–61.PubMedCrossRefGoogle Scholar
  14. 14.
    Neustock P, Brand JM, Kruse A, Kircher H. Cytokine production of the human monocytic cell line Mono Mac 6 in comparison to mature monocytes in peripheral blood mononuclear cells. Immunobiology. 1993;188:293–302.PubMedGoogle Scholar
  15. 15.
    Schägger H, von Jagow G. Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis for the separation of proteins in the range from 1 to 100 kDa. Anal Biochem. 1987;166:368–79.PubMedCrossRefGoogle Scholar
  16. 16.
    Towbin H, Staehelin T, Gordon J. Electrophoretic transfer of proteins from polyacrylamide gel to nitrocellulose sheet: procedure and some applications. Proc Natl Acad Sci USA. 1979;76:4350–4.PubMedCrossRefGoogle Scholar
  17. 17.
    Creminon C, Habib A, Maclouf J, Pradelles P, Grassi J, Frobert Y. Differential measurement of constitutive (COX-1) and inducible (COX-2) cyclooxygenase expression in human umbilical vein endothelial cells using specific immunometric enzyme immunoassays. Biochim Biophys Acta. 1995;1254:341–8.PubMedGoogle Scholar
  18. 18.
    Creminon C, Frobert Y, Habib A, Maclouf J, Pradelles P, Grassi J. Immunological studies of human constitutive cyclooxygenase (COX-1) using enzyme immunometric assay. Biochim Biophys Acta. 1995;1254:333–40.PubMedGoogle Scholar
  19. 19.
    Hla T, Neilson K. Human cyclooxygenase-2 cDNA. Proc Natl Acad Sci USA. 1992;89:7384–8.PubMedCrossRefGoogle Scholar
  20. 20.
    MacDonald RJ, Swift GH, Przybyla AE, Chirgwin JM. Isolation of RNA using guanidinium salts. Meth Enzymol. 1987;152:219–27.PubMedGoogle Scholar
  21. 21.
    Mitchell JA, Akarasereenont P, Thiermermann C, Flower R, Vane JR. Selectivity of nonsteroidal antiinflammatory drugs as inhibitors of constitutive and inducible cyclooxygenase. Proc Natl Acad Sci USA. 1994;90:11693–7.CrossRefGoogle Scholar
  22. 22.
    Meager A. RIA, IRMA, and ELISA assays for cytokines. In: Bakwill FR, ed. Cytokines, A Practical Approach. Oxford: IRL Press at Oxford University Press; 1991:304–7.Google Scholar
  23. 23.
    Otto JC, DeWitt DL, Smith WL.N-Glycosilation of prostaglandin endoperoxide synthases-1 and-2 and their orientations in the endoplasmic reticulum. J Biol Chem. 1993;268:18234–42.PubMedGoogle Scholar
  24. 24.
    Hempel SL, Monick MM, Hunninghake GW. Lipopolysaccharide induces prostaglandin H synthase-2 protein and mRNA in human alveolar macrophages and blood monocytes. J Clin Invest. 1994;93:391–6.PubMedCrossRefGoogle Scholar
  25. 25.
    Vane JR, Botting RM. New insights into the mode of action of anti-inflammatory drugs. Inflamm Res. 1995;44:1–10.PubMedCrossRefGoogle Scholar
  26. 26.
    Ziegler-Heitbrock HWL, Schraut W, Wendelgass P et al. Distinct patterns of differentiation induced in the monocytic cell line MONO MAC 6. J Leukocyte Biol. 1994;55:73–80.PubMedGoogle Scholar
  27. 27.
    Kunkel SL, Chensue SW. Arachidonic acid metabolites regulate interleukin-1 production. Biochem Biophys Res Commun. 1985;128:892–7.PubMedCrossRefGoogle Scholar

Copyright information

© Kluwer Academic Publishers 1996

Authors and Affiliations

  • T. Christoph
    • 1
  • M. Widerna
    • 1
  • A. Bodenteich
    • 1
  • J. Berg
    • 1
  1. 1.Immunology and Molecular Biology Unit, Department of PharmacologyNycomed PharmaLinzAustria

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