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. 2003 Apr;12(2):71–78. doi: 10.1080/0962935031000097673

Dexamethasone prevents granulocyte-macrophage colony-stimulating factor-induced nuclear factor-kappaB activation, inducible nitric oxide synthase expression and nitric oxide production in a skin dendritic cell line.

Ana Luísa Vital 1, Margarida Gonçalo 1, M Teresa Cruz 1, Américo Figueiredo 1, Carlos B Duarte 1, M Celeste Lopes 1
PMCID: PMC1781603  PMID: 12775356

Abstract

AIMS: Nitric oxide (NO) has been increasingly implicated in inflammatory skin diseases, namely in allergic contact dermatitis. In this work, we investigated the effect of dexamethasone on NO production induced by the epidermal cytokine granulocyte-macrophage colony-stimulating factor (GM-CSF) in a mouse fetal skin dendritic cell line. METHODS: NO production was assessed by the method of Griess. Expression of the inducible isoform of nitric oxide synthase (iNOS) protein was evaluated by western blot analysis and immunofluorescence microscopy. Western blot analysis was also performed to evaluate cytosolic IkappaB-alpha (IkappaB-alpha) protein levels. The electrophoretic mobility shift assay was used to evaluate the activation or inhibition of nuclear factor kappa B (NF-kappaB). RESULTS: GM-CSF induced iNOS expression and NO production, and activated the transcription factor NF-kappaB. Dexamethasone inhibited, in a dose-dependent manner, NO production induced by GM-CSF. Addition of dexamethasone to the culture, 30 min before GM-CSF stimulation, significantly inhibited the cellular expression of iNOS. Dexamethasone also inhibited GM-CSF-induced NF-kappaB activation by preventing a significant decrease on the IkappaB-alpha protein levels, thus blocking NF-kappaB migration to the nucleus. CONCLUSIONS: The corticosteroid dexamethasone inhibits GM-CSF-induced NF-kappaB activation, iNOS protein expression and NO production. These results suggest that dexamethasone is a potent inhibitor of intracellular events that are involved on NO synthesis, in skin dendritic cells.

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Selected References

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  1. Almawi W. Y., Melemedjian O. K. Negative regulation of nuclear factor-kappaB activation and function by glucocorticoids. J Mol Endocrinol. 2002 Apr;28(2):69–78. doi: 10.1677/jme.0.0280069. [DOI] [PubMed] [Google Scholar]
  2. Auphan N., DiDonato J. A., Rosette C., Helmberg A., Karin M. Immunosuppression by glucocorticoids: inhibition of NF-kappa B activity through induction of I kappa B synthesis. Science. 1995 Oct 13;270(5234):286–290. doi: 10.1126/science.270.5234.286. [DOI] [PubMed] [Google Scholar]
  3. Baldwin A. S., Jr The NF-kappa B and I kappa B proteins: new discoveries and insights. Annu Rev Immunol. 1996;14:649–683. doi: 10.1146/annurev.immunol.14.1.649. [DOI] [PubMed] [Google Scholar]
  4. Bonham C. A., Lu L., Hoffman R. A., Simmons R. L., Thomson A. W. Nitric oxide production by dendritic cells is associated with impairment of T cell responses. Transplant Proc. 1997 Feb-Mar;29(1-2):1116–1117. doi: 10.1016/s0041-1345(96)00458-7. [DOI] [PubMed] [Google Scholar]
  5. Bonham C. A., Lu L., Li Y., Hoffman R. A., Simmons R. L., Thomson A. W. Nitric oxide production by mouse bone marrow-derived dendritic cells: implications for the regulation of allogeneic T cell responses. Transplantation. 1996 Dec 27;62(12):1871–1877. doi: 10.1097/00007890-199612270-00033. [DOI] [PubMed] [Google Scholar]
  6. Bruch-Gerharz D., Ruzicka T., Kolb-Bachofen V. Nitric oxide in human skin: current status and future prospects. J Invest Dermatol. 1998 Jan;110(1):1–7. doi: 10.1046/j.1523-1747.1998.00084.x. [DOI] [PubMed] [Google Scholar]
  7. Cato A. C., Wade E. Molecular mechanisms of anti-inflammatory action of glucocorticoids. Bioessays. 1996 May;18(5):371–378. doi: 10.1002/bies.950180507. [DOI] [PubMed] [Google Scholar]
  8. Chen F. E., Ghosh G. Regulation of DNA binding by Rel/NF-kappaB transcription factors: structural views. Oncogene. 1999 Nov 22;18(49):6845–6852. doi: 10.1038/sj.onc.1203224. [DOI] [PubMed] [Google Scholar]
  9. Cruz M. T., Duarte C. B., Gonçalo M., Carvalho A. P., Lopes M. C. Involvement of JAK2 and MAPK on type II nitric oxide synthase expression in skin-derived dendritic cells. Am J Physiol. 1999 Dec;277(6 Pt 1):C1050–C1057. doi: 10.1152/ajpcell.1999.277.6.C1050. [DOI] [PubMed] [Google Scholar]
  10. Cruz M. T., Duarte C. B., Gonçalo M., Carvalho A. P., Lopes M. C. LPS induction of I kappa B-alpha degradation and iNOS expression in a skin dendritic cell line is prevented by the janus kinase 2 inhibitor, Tyrphostin b42. Nitric Oxide. 2001 Feb;5(1):53–61. doi: 10.1006/niox.2000.0320. [DOI] [PubMed] [Google Scholar]
  11. Cruz M. T., Duarte C. B., Gonçalo M., Figueiredo A., Carvalho A. P., Lopes M. C. Granulocyte-macrophage colony-stimulating factor activates the transcription of nuclear factor kappa B and induces the expression of nitric oxide synthase in a skin dendritic cell line. Immunol Cell Biol. 2001 Dec;79(6):590–596. doi: 10.1046/j.1440-1711.2001.01041.x. [DOI] [PubMed] [Google Scholar]
  12. Cyert M. S. Regulation of nuclear localization during signaling. J Biol Chem. 2001 Apr 12;276(24):20805–20808. doi: 10.1074/jbc.R100012200. [DOI] [PubMed] [Google Scholar]
  13. De Vera M. E., Taylor B. S., Wang Q., Shapiro R. A., Billiar T. R., Geller D. A. Dexamethasone suppresses iNOS gene expression by upregulating I-kappa B alpha and inhibiting NF-kappa B. Am J Physiol. 1997 Dec;273(6 Pt 1):G1290–G1296. doi: 10.1152/ajpgi.1997.273.6.G1290. [DOI] [PubMed] [Google Scholar]
  14. Ghosh S., May M. J., Kopp E. B. NF-kappa B and Rel proteins: evolutionarily conserved mediators of immune responses. Annu Rev Immunol. 1998;16:225–260. doi: 10.1146/annurev.immunol.16.1.225. [DOI] [PubMed] [Google Scholar]
  15. Girolomoni G., Lutz M. B., Pastore S., Assmann C. U., Cavani A., Ricciardi-Castagnoli P. Establishment of a cell line with features of early dendritic cell precursors from fetal mouse skin. Eur J Immunol. 1995 Aug;25(8):2163–2169. doi: 10.1002/eji.1830250807. [DOI] [PubMed] [Google Scholar]
  16. Kimber I., Holliday M. R., Dearman R. J. Cytokine regulation of chemical sensitization. Toxicol Lett. 1995 Dec;82-83:491–496. doi: 10.1016/0378-4274(95)03497-8. [DOI] [PubMed] [Google Scholar]
  17. Lu L., Bonham C. A., Chambers F. G., Watkins S. C., Hoffman R. A., Simmons R. L., Thomson A. W. Induction of nitric oxide synthase in mouse dendritic cells by IFN-gamma, endotoxin, and interaction with allogeneic T cells: nitric oxide production is associated with dendritic cell apoptosis. J Immunol. 1996 Oct 15;157(8):3577–3586. [PubMed] [Google Scholar]
  18. Morhenn V. B. Langerhans cells may trigger the psoriatic disease process via production of nitric oxide. Immunol Today. 1997 Sep;18(9):433–436. doi: 10.1016/s0167-5699(97)01116-x. [DOI] [PubMed] [Google Scholar]
  19. Morita H., Semma M., Hori M., Kitano Y. Clinical application of nitric oxide synthase inhibitor for atopic dermatitis. Int J Dermatol. 1995 Apr;34(4):294–295. doi: 10.1111/j.1365-4362.1995.tb01602.x. [DOI] [PubMed] [Google Scholar]
  20. Moser M., De Smedt T., Sornasse T., Tielemans F., Chentoufi A. A., Muraille E., Van Mechelen M., Urbain J., Leo O. Glucocorticoids down-regulate dendritic cell function in vitro and in vivo. Eur J Immunol. 1995 Oct;25(10):2818–2824. doi: 10.1002/eji.1830251016. [DOI] [PubMed] [Google Scholar]
  21. Ormerod A. D., Copeland P., Shah S. A. Treatment of psoriasis with topical NG-monomethyl-L-arginine, an inhibitor of nitric oxide synthesis. Br J Dermatol. 2000 May;142(5):985–990. doi: 10.1046/j.1365-2133.2000.03483.x. [DOI] [PubMed] [Google Scholar]
  22. Ormerod A. D., Dwyer C. M., Reid A., Copeland P., Thompson W. D. Inducible nitric oxide synthase demonstrated in allergic and irritant contact dermatitis. Acta Derm Venereol. 1997 Nov;77(6):436–440. doi: 10.2340/0001555577436440. [DOI] [PubMed] [Google Scholar]
  23. Ormerod A. D., Weller R., Copeland P., Benjamin N., Ralston S. H., Grabowksi P., Herriot R. Detection of nitric oxide and nitric oxide synthases in psoriasis. Arch Dermatol Res. 1998 Jan-Feb;290(1-2):3–8. doi: 10.1007/s004030050268. [DOI] [PubMed] [Google Scholar]
  24. Pahl H. L. Activators and target genes of Rel/NF-kappaB transcription factors. Oncogene. 1999 Nov 22;18(49):6853–6866. doi: 10.1038/sj.onc.1203239. [DOI] [PubMed] [Google Scholar]
  25. Pan J., Ju D., Wang Q., Zhang M., Xia D., Zhang L., Yu H., Cao X. Dexamethasone inhibits the antigen presentation of dendritic cells in MHC class II pathway. Immunol Lett. 2001 Apr 2;76(3):153–161. doi: 10.1016/s0165-2478(01)00183-3. [DOI] [PubMed] [Google Scholar]
  26. Piemonti L., Monti P., Allavena P., Sironi M., Soldini L., Leone B. E., Socci C., Di Carlo V. Glucocorticoids affect human dendritic cell differentiation and maturation. J Immunol. 1999 Jun 1;162(11):6473–6481. [PubMed] [Google Scholar]
  27. Qureshi A. A., Hosoi J., Xu S., Takashima A., Granstein R. D., Lerner E. A. Langerhans cells express inducible nitric oxide synthase and produce nitric oxide. J Invest Dermatol. 1996 Dec;107(6):815–821. doi: 10.1111/1523-1747.ep12330572. [DOI] [PubMed] [Google Scholar]
  28. Ross R., Gillitzer C., Kleinz R., Schwing J., Kleinert H., Förstermann U., Reske-Kunz A. B. Involvement of NO in contact hypersensitivity. Int Immunol. 1998 Jan;10(1):61–69. doi: 10.1093/intimm/10.1.61. [DOI] [PubMed] [Google Scholar]
  29. Ross R., Reske-Kunz A. B. The role of NO in contact hypersensitivity. Int Immunopharmacol. 2001 Aug;1(8):1469–1478. doi: 10.1016/s1567-5769(01)00091-1. [DOI] [PubMed] [Google Scholar]
  30. Rowe A., Farrell A. M., Bunker C. B. Constitutive endothelial and inducible nitric oxide synthase in inflammatory dermatoses. Br J Dermatol. 1997 Jan;136(1):18–23. [PubMed] [Google Scholar]
  31. Sahin S., Onder M., Sancak B., Bukan N., Gürer M. A. The role of nitric oxide in allergic contact dermatitis. Arch Dermatol Res. 2001 Apr;293(4):214–217. doi: 10.1007/s004030100207. [DOI] [PubMed] [Google Scholar]
  32. Taniuchi S., Kojima T., Hara Mt K., Yamamoto A., Sasai M., Takahashi H., Kobayashi Y. Increased serum nitrate levels in infants with atopic dermatitis. Allergy. 2001 Jul;56(7):693–695. doi: 10.1034/j.1398-9995.2001.00131.x. [DOI] [PubMed] [Google Scholar]
  33. Thurnher M., Zelle-Rieser C., Ramoner R., Bartsch G., Höltl L. The disabled dendritic cell. FASEB J. 2001 Apr;15(6):1054–1061. doi: 10.1096/fj.00-0508hyp. [DOI] [PubMed] [Google Scholar]
  34. Wallengren J., Larsson B. Nitric oxide participates in prick test and irritant patch test reactions in human skin. Arch Dermatol Res. 2001 Mar;293(3):121–125. doi: 10.1007/s004030000198. [DOI] [PubMed] [Google Scholar]
  35. Weller R. Nitric oxide, skin growth and differentiation: more questions than answers? Clin Exp Dermatol. 1999 Sep;24(5):388–391. doi: 10.1046/j.1365-2230.1999.00509.x. [DOI] [PubMed] [Google Scholar]
  36. Weller R. Nitric oxide--a newly discovered chemical transmitter in human skin. Br J Dermatol. 1997 Nov;137(5):665–672. [PubMed] [Google Scholar]
  37. Wissink S., van Heerde E. C., vand der Burg B., van der Saag P. T. A dual mechanism mediates repression of NF-kappaB activity by glucocorticoids. Mol Endocrinol. 1998 Mar;12(3):355–363. doi: 10.1210/mend.12.3.0081. [DOI] [PubMed] [Google Scholar]
  38. Witmer-Pack M. D., Olivier W., Valinsky J., Schuler G., Steinman R. M. Granulocyte/macrophage colony-stimulating factor is essential for the viability and function of cultured murine epidermal Langerhans cells. J Exp Med. 1987 Nov 1;166(5):1484–1498. doi: 10.1084/jem.166.5.1484. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Xie Q. W., Kashiwabara Y., Nathan C. Role of transcription factor NF-kappa B/Rel in induction of nitric oxide synthase. J Biol Chem. 1994 Feb 18;269(7):4705–4708. [PubMed] [Google Scholar]
  40. Yamamoto Y., Gaynor R. B. Therapeutic potential of inhibition of the NF-kappaB pathway in the treatment of inflammation and cancer. J Clin Invest. 2001 Jan;107(2):135–142. doi: 10.1172/JCI11914. [DOI] [PMC free article] [PubMed] [Google Scholar]

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