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Archives of Dermatological Research

, Volume 284, Issue 6, pp 324–332 | Cite as

Interleukin-6 in psoriasis: expression and mitogenicity studies

  • J. T. Elder
  • C. I. Sartor
  • D. K. Boman
  • S. Benrazavi
  • G. J. Fisher
  • M. R. Pittelkow
Original Contributions

Summary

InterIeukin-6 (IL-6) is a multifunctional cytokine which has been suggested to function as an autocrine mitogen in psoriatic epidermis. We report here the results of several experiments designed to further examine this hypothesis. Blot hybridization was unable to detect 1.3 kb IL-6 transcripts in RNA extracted from normal or psoriatic epidermal (keratome) biopsies, suggesting that IL-6 expression is very low in normal and psoriatic epidermis. Therefore, qualitative and semiquantitative PCR/Southern blot analyses were performed on keratome-derived RNA, and revealed variable but significantly increased IL-6 mRNA levels in lesional psoriatic relative to normal tissue. To further examine the ability of normal human keratinocytes (NHK) to express IL-6, RNA was extracted from rapidly proliferating secondary NHK cultures. IL-6 transcripts were nearly undetectable by blotting in keratinocytes grown in low-calcium serum-free medium, but low levels could be induced by treatment with 1.8 mM CaCl2. IL-6 transcripts were strongly superinduced after cycloheximide treatment, suggesting that a labile protein regulates IL-6 mRNA levels in these cells. Finally, the mitogenic activity of IL-6 was examined in NHK under varying conditions of cell density and added growth factors. IL-6 did not stimulate high density keratinocyte growth in the presence or absence of other growth factors, but did stimulate clonal growth in epidermal growth factor (EGF)-deficient media at high concentrations (≧10 ng/ml). The proliferative effects of IL-6, but not of basic fibroblast growth factor, were abrogated by monoclonal antibodies directed against the EGF receptor. Taken together, these results suggest that the proliferative effects of IL-6 are mediated indirectly via the EGF/TGF-α receptor, and that autocrine overexpression of IL-6 may be limited in psoriatic keratinocytes.

Key words

Interleukin-6 Psoriasis Polymerase chain reaction Keratinocytes Cell proliferation 

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References

  1. 1.
    Aarden LA, De Groot ER, Schaap OL, Lansdorp PM (1987) Production of hybridoma growth factor by human monocytes. Eur J Immunol 17: 1411–1416Google Scholar
  2. 2.
    Beutler BA, Milsark IW, Cerami A (1985) Cachectin/tumor necrosis factor: production, distribution, and metabolic fate in vivo. J Immunol 135: 3972–3977Google Scholar
  3. 3.
    Birnboim HC, Doly J (1979) A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res 7: 1513Google Scholar
  4. 4.
    Bos JD, Hulsebosch HJ, Krieg SR, Bakker PM, Cormane RH (1983) Immunocompetent cells in psoriasis: in situ immunophenotyping by monoclonal antibodies. Arch Dermatol Res 275: 181–189Google Scholar
  5. 5.
    Briggaman RA (1982) Epidermal-dermal interactions in adult skin. J Invest Dermatol 79: 21S-24SGoogle Scholar
  6. 6.
    Caput D, Beutler B, Hartog K, Thayer R, Brown-Shimer S, Cerami A (1986) Identification of a common nucleotide sequence in the 3′untranslated region of mRNA molecules specifying inflammatory mediators. Proc Natl Acad Sci USA 83: 1670–1674Google Scholar
  7. 7.
    Castell J, Klapproth J, Gross V, Walter E, Andus T, Snyers L, Content J, Heinrich RC (1990) Fate of interleukin-6 in the rat: involvement of skin of its catabolism. Eur J Biochem 189: 113–118Google Scholar
  8. 8.
    Cook PW, Pittelkow MR, Shipley GD (1991) Growth factor-independent proliferation of normal human neonatal keratinocytes: production of autocrine- and paracrine-acting mitogenic factors. J Cell Physiol 146: 277–289Google Scholar
  9. 9.
    Derynck R, Roberts AOB, Winkler M, Chen EQ, Goeddel DV (1984) Human transforming growth factor-α: precursor structure and expression in E. coli. Cell 38: 287–297Google Scholar
  10. 10.
    Elder JT, Fisher GJ, Lindquist PB, Bennett GL, Pittelkow MR, Coffey RJ Jr., Ellingsworth L, Derynck R, Voorhees JJ (1989) Overexpression of transforming growth factor α in psoriatic epidermis. Science 243: 811–814Google Scholar
  11. 11.
    Elder JT, Tavakkol A, Klein SB, Zeigler ME, Wicha M, Voorhees JJ (1990) Protooncogene expression in normal and psoriatic skin. J Invest Dermatol 94: 19–25Google Scholar
  12. 12.
    Elder JT, Fisher GJ, Zhang Q-Y, Eisen D, Krust A, Kastner P, Chambon P, Voorhees JJ (1991) Retinoic acid receptor gene expression in human skin. J Invest Dermatol 96: 425–433Google Scholar
  13. 13.
    Elder JT, Strom A, Pettersson U, Tavakkol A, Griffiths CEM, Krust A, Kastner P, Chambon P, Voorhees JJ (1992) Differential regulation of retinoid acid receptors and binding proteins in human skin. J Invest Dermatol 98: 673–679Google Scholar
  14. 14.
    Elias JA, Lentz V (1990) IL-1 and tumor necrosis factor synergistically stimulate fibroblast IL-6 production and stabilize IL-6 messenger RNA. J Immunol 145: 161–166Google Scholar
  15. 15.
    Feinberg AP, Vogelstein B (1983) A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem 132: 6–13Google Scholar
  16. 16.
    Fraud JE, Briggaman RA, Lazarus GS (1982) Uninvolved skin from psoriatic patients develops signs of involved psoriatic skin after being grafted onto nude mice. Science 215: 685–687Google Scholar
  17. 17.
    Gearing AJH, Fincham NJ, Bird CR, Wadhwa M, Meager A, Cartwright JE, Camp RDR (1990) Cytokines in skin lesions of psoriasis. Cytokine 2: 68–75Google Scholar
  18. 18.
    Gillitzer R, Berger R, Mielke V, Muller C, Wolff K, Stingl G (1991) Upper keratinocytes of psoriatic skin lesions express high levels of NAP-1/IL-8 mRNA in situ. J Invest Dermatol 97: 73–79Google Scholar
  19. 19.
    Grossman RM, Krueger J, Yourish D, Granelli-Piperno A, Murphy DP, May LT, Kupper TS, Sehgal PB, Gottlieb AB (1989) Interleukin 6 is expressed in high levels in psoriatic skin and stimulates proliferation of cultured human keratinocytes. Proc Natl Acad Sci USA 86: 6367–6371Google Scholar
  20. 20.
    Hirano T, Akira S, Taga T, Kishimoto T (1990) Biological and clinical aspects of interleukin 6. Immunol Today 11: 443–449Google Scholar
  21. 21.
    Kawasaki ES (1990) Amplification of RNA. In: Innis MA, Gelfand DH, Sninsky JJ (eds) PCR protocols: a guide to methods and applications. Academic Press, New York, pp 21–27Google Scholar
  22. 22.
    Kirnbauer R, Kock A, Schwarz T, Urbanski A, Krutmann J, Borth W, Damm D, Shiplye G, Ansel JC, Luger TA (1989) IFN-Β2, B cell differentiation factor 2, or hybridoma growth factor (IL-6) is expressed and released by human epiderma cells and epidermoid carcinoma cell lines. J Immunol 142: 1922–1928Google Scholar
  23. 23.
    Kishimoto T (1989) The biology of interleukin-6. Blood 74: 10Google Scholar
  24. 24.
    Kreuger GG, Chambers DA, Shelby J (1981) Involved and uninvolved skin from psoriatic subjects: are they equally diseased? Assessment by skin transplanted to congenitally athymic (nude) mice. J Clin Invest 68: 1548–1557Google Scholar
  25. 25.
    Krueger J, Ray A, Tamm I, Sehgal PB (1991) Expression and function of interleukin-6 in epithelial cells. J Cell Biochem 45: 327–334Google Scholar
  26. 26.
    Kupper T, Min K, Sehgal P, Mizutani H, Birchall N, Ray A (1989) Production of IL-6 by keratinocytes: implications for epidermal inflammation and immunity. Ann NY Acad Sci 557: 454–465Google Scholar
  27. 27.
    Loppnow H, Libby P (1990) Proliferating or interleukin 1-activated human vascular smooth muscle cells secrete copious interleukin 6. J Clin Invest 85: 731–738Google Scholar
  28. 28.
    May LT, Helfgott DC, Sehgal PB (1986) Anti-Β-interferon antibodies inhibit the increased expression of HLA-B7 mRNA in tumor necrosis factor-treated human fibroblasts: structural studies of the Β2-interferon involved. Proc Natl Acad Sci USA 83: 8957–8961Google Scholar
  29. 29.
    May LT, Ghrayeb J, Santhanam U, Tatter SB, Sthoeger Z, Helgott DC, Chiorazzi N, Grieninger G, Sehgal PB (1988) synthesis and secretion of multiple forms of Β2-interferon/B-cell differentiation factor 2/hepatocyte-stimulating factor by human fibroblasts and monocytes. J Biol Chem 263: 7760–7766Google Scholar
  30. 30.
    Neuner P, Urbanski A, Trautinger F, Moller A, Kirnbauer R, Kapp A, Schopf E, Schwarz T, Luger TA (1991) Increased IL-6 production by monocytes and keratinocytes in patients with psoriasis. J Invest Dermatol 97: 27–33Google Scholar
  31. 31.
    Ohta Y, Katayama I, Funato T, Yokozeki H, Nishiyama S, Hirano T, Kishimoto T, Nishioka K (1991) In situ expression of messenger RNA of interleukin-1 and interleukin-6 in psoriasis: interleukin-6 involved in formation of psoriatic lesions. Arch Dermatol Res 283: 351–356Google Scholar
  32. 32.
    Oxholm A, Oxholm P, Staberg B, Bendtzen K (1989) Expression of interleukin-6-like molecules and tumour necrosis factor after topical treatment of psoriasis with a new vitamin D analogue (MC903). Acta Derm Venereol (Stockh) 69: 385–390Google Scholar
  33. 33.
    Oxholm A, Oxholm P, Staberg B, Bendtzen K (1989) Interleukin-6 in the epidermis of patients with psoriasis before and during PUVA treatment. Acta Derm Venereol (Stockh) 69: 195–199Google Scholar
  34. 34.
    Oxhom A, Diamant M, Oxhom P, Bendtzen K (1991) Interleukin-6 and tumor necrosis factor alpha are expressed by keratinocytes but not by Langerhans cells. APMIS 99: 58–64Google Scholar
  35. 35.
    Partridge M, Chantry D, Turner M, Feldmann M (1991) Production of interleukin-1 and interleukin-6 by human keratinocytes and squamous cell lines. J Invest Dermatol 96: 771–776Google Scholar
  36. 36.
    Pittelkow MR, Scott RE (1986) New techniques for the in vitro culture of human skin keratinocytes and perspectives on their use for grafting of patients with extensive burns. Mayo Clin Proc 61: 771–777Google Scholar
  37. 37.
    Prens EP, Benne K, van Damma J, Bakkus M, Brakel K, Benner R, van Joost T (1990) Interleukin-1 and interleukin-6 inpsoriasis. J Invest Dermatol 95: 121S-124SGoogle Scholar
  38. 38.
    Rabilloud T, Asselineau D, Darmon M (1989) Presence of serum albumin in normal human epidermis: Possible implications for the nutrition and physiology of stratified epithelia. Mol Biol Rep 13: 213–219Google Scholar
  39. 39.
    Ray A, Sassone-Corsi P, Sehgal PB (1989) A multiple cytokine and second messenger-responsive element in the enhancer of the human interleukin-6 gene: similarities with c-fos gene regulation. Mol Cell Biol 9: 5537–5547Google Scholar
  40. 40.
    Ristow HJ (1990) Interleukin-1 does not stimulate DNA synthesis of cultured human keratinocytes growth-arrested in growth-factor-depleted medium. J Invest Dermatol 95: 688–692Google Scholar
  41. 41.
    Roth MS, Antin JH, Bingham EL, Ginsburg D (1989) Detection of Philadelphia chromosome-positive cells by the polymerase chain reaction following bone marrow transplant for chronic myelogeneous leukemia. Blood 74: 882–885Google Scholar
  42. 42.
    Sambrook J, Fritsch EF, Maniatis T (1989) Extraction, purification, and analysis of messenger RNA from eukaryotic cells. In: Sambrook J, Fritsch EF, Maniatis T (eds) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, Section 7.2Google Scholar
  43. 43.
    Shaw G, Kamen R (1986) A conserved AU sequence from the 3′ untranslated region of GM-CSF mRNA mediates selective mRNA degradation. Cell 46: 659–667Google Scholar
  44. 44.
    Shipley GD, Keeble WW, Hendrickson JE, Coffey RJ, Pittelkow MR (1989) Growth of normal human keratinocytes and fibroblasts in serum-free medium is stimulated by acidic and basic fibroblast growth factor. J Cell Physiol 138: 511–518Google Scholar
  45. 45.
    Urbanski A, Schwarz T, Neuner P, Krutmann J, Kirnbauer R, Kock A, Luger TA (1990) Ultraviolet light induces increrasedcirculating interleukin-6 in humans. J Invest Dermatol 94: 808–811Google Scholar
  46. 46.
    Voorhees JJ, Duell EA, Bass LJ, Powell JA, Harreil ER (1972) Decreased cyclic AMP in the epidermis of lesions of psoriasis. Arch Dermatol 105: 701Google Scholar
  47. 47.
    Weinstein GD, McCullough JL, Ross PA (1985) Cell kinetic basis for pathophysiology of psoriasis. J Invest Dermatol 85: 579–583Google Scholar
  48. 48.
    Wille JJ Jr, Pittelkow MR, Shipley GD, Scott RE (1984) Integrated control of growth and differentiation of normal human prokeratinocytes cultured in serum-free medium: clonal analyses, growth kinetics, and cell cycle studies. J Cell Physiol 121: 31–44Google Scholar
  49. 49.
    Yasukawa K, Hirano T, Watanabe Y, Muratani K, Matsuda T, Nakai S, Kishimoto T (1987) Structure and expression ofhuman B cell stimulatory factor-2 (BSF-2/IL-6) gene. Embo J 6: 2938–2945Google Scholar
  50. 50.
    Yoshizaki K, Nishimoto N, Matsumoto K, Tagoh H, Taga T, Deguchi Y, Kuritani T, Hirano T, Hashimoto K, Okada N, Kishimoto T (1990) Interleukin 6 and expression of its receptor on epidermal keratinocytes. Cytokine 2: 381–387Google Scholar

Copyright information

© Springer-Verlag 1992

Authors and Affiliations

  • J. T. Elder
    • 1
  • C. I. Sartor
    • 2
  • D. K. Boman
    • 1
  • S. Benrazavi
    • 1
  • G. J. Fisher
    • 1
  • M. R. Pittelkow
    • 3
  1. 1.Department of DermatologyUniversity of MichiganAnn ArborUSA
  2. 2.Department of Internal Mediaine, C560A MSRB IIUniversity of MichiganAnn ArborUSA
  3. 3.Department of DermatologyMayo ClinicRochesterUSA

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