Original Article

Journal of Investigative Dermatology (1995) 104, 922–927; doi:10.1111/1523-1747.ep12606202

Ultraviolet-B-Induced Apoptosis of Keratinocytes: Evidence for Partial Involvement of Tumor Necrosis Factor-alpha in the Formation of Sunburn Cells

Agatha Schwarz, Ranjit Bhardwaj, Yoshinori Aragane, Karsten Mahnke, Helge Riemann, Dieter Metze, Thomas A Luger and Thomas Schwarz

Ludwig Boltzmann Institute for Cell Biology and Immunobiology of the Skin, Department of Dermatology, University Münster, Münster, Germany

Received 28 June 1994; Revised 19 December 1994; Accepted 5 January 1995.

Top

Abstract

Irradiation with ultraviolet (UV) B radiation results in the formation of apoptotic keratinocytes called sunburn cells. Recently, it was demonstrated that keratinocytes can release tumor necrosis factor-alpha (TNF-alpha), which is known to cause apoptosis in particular cells. In addition, it has been shown that UVB light induces the release of TNF-alpha by keratinocytes and that keratinocytes express the 55-kD receptor for TNF-alpha. Therefore, we investigated whether TNF-alpha is involved in UV-induced apoptosis of kerationocytes. Normal human keratinocytes and HaCaT cells were exposed to UVB light, and apoptosis was examined by nick translation evaluated by fluorescence-activated cell sorter analysis. UVP induced apoptosis in a dose-dependent manner, which was confirmed by electron microscopy. Addition of a polyclonal antibody directed against human TNF-alpha immediately after UVB exposure was able to reduce DNA fragmentation. However, it was not possible to rescue all cells from apoptosis. To prove whether TNF-alpha is also involved in vivo in UVB-induced apoptosis of keratinocytes, Balb/c mice were exposed to UVB on their abdomens, skin biopsies were performed 24 h later, and sunburn cells were counted. A single dose of 2000J/M2 caused a significant induction of sunburn cells. Subcutaneous injection of a polyclonal antibody directed against murine TNF-alpha immediately after UVB treatment resulted in a significant but incomplete reduction of sunburn cells, whereas injection of a rabbit IgG as a control had no effect. In both the in vitro and in vivo systems, application of recombinant TNF-alpha alone either to untreated keratinocytes or into normal murine skin did not induce sunburn cells. Thus, these data demonstrate that TNF-alpha is involved in UVB-induced apoptosis, but by itself is not able to induce sunburn cells. This further supports the notion that UVB-induced apoptosis of keratinocytes is a multifactorial event.

Top

References

  1. Kerr, JFR, Harmon, BV: Definition and incidence of apoptosis: an historical perspective. In: Tomei LD, Cope FO (eds.). Apoptosis: The Molecular Basis of Cell Death. Current Communication in Cell and Molecular Biology Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, 1991
  2. Schwatrz, LM, Osborne, BA: Programmed cell death, apoptosis and killer genes. Immunol Today 14: 582–590, 1993
  3. Cohen, JJ: Apoptosis. Immunol Today 14: 126–130, 1993 | Article | PubMed | ISI | ChemPort |
  4. Danno, K, Horio, T: Sunburn cell: factors involved in its formation. Photochem Photobiol 45: 683–690, 1987 | PubMed | ISI | ChemPort |
  5. Young, AR: The sunburn cell. Photodermatology 4: 127–134, 1987 | PubMed | ISI | ChemPort |
  6. Martin, SJ, Cotter, TG: Ultraviolet B irradiation of human leukaemia HL-60 cells in vitro induces apoptosis. Int J Radial Biol 59: 1001–1016, 1991
  7. Casciola-Rosen, LA, Anhalt, G, Rosen, A: Autoantigens targeted in systemic lupus erythematosus are clustered in two populations of surface structures on apoptotic keratinocytes. J Exp Med 179: 1317–1330, 1994 | Article | PubMed | ChemPort |
  8. Carswell, EA, Old, LJ, Kassel, RL, Green, S, Fiore, N, Williamson, B: An endotoxin-induced serum factor that causes necrosis of tumors. Proc Natl Acad Sci USA 72: 3666–3670, 1975 | PubMed | ChemPort |
  9. Beutler, B, Cerami, A: Cachectin (tumor necrosis factor): an endogenous mediator of shock and inflammatory response. In: Oppenheim JJ, Shevach EM (eds.). Immunophysiology. The Role of Cells and Cytokins in Immunity an Inflammation Oxford University Press, New York, pp 226–237, 1990
  10. Köck, A, Schwarz, T, Kirnbauer, R, Urbanski, A, Perry, P, Ansel, JC, Luger, TA: Human keratinocytes are a source for tumor necrosis factor alpha. Evidence for synthesis and release upon stimulation with endotoxin or ultraviolet light. J Exp Med 172: 1609–1614, 1990 | Article | PubMed | ISI | ChemPort |
  11. Trefzer, U, Brockhaus, M, Lötscher, H, Parlow, F, Budnik, A, Grewe, M, Christoph, H, Kapp, A, Schöpf, E, Luger, TA, Krutmann, J: The 55-kD tumor necrosis factor receptor on human keratinocytes is regulated by tumor necrosis factor-alpha and by ultraviolet B radiation. J Clin Invst 92: 462–470, 1993
  12. Elias, E, Moore, PB, Rose, SM: Tumor necrosis factor induced DNA fragmentation in HL-60 cells. Biochem Biophys Res Commun 157: 963–969, 1988
  13. Greenblatt, MS, Elias, L: The type B receptor for tumor necrosis factor-alpha mediates DNA fragmentation in HL-60 and U937 cells and differentiation in HL-60 cells. Blood 80: 1339–1346, 1992
  14. Wright, SC, Kumar, P, Tam, AW, Shen, N, Varma, M, Larrick, JW: Apoptosis and DNA fragmentation precede TNF-induced cytolysis in U937 cells. J Cell Biochem 48: 344–355, 1992 | PubMed |
  15. Kyprianou, N, Alexander, RB, Isaacs, JT: Activation of programmed cell death by recombinant human tumor necrosis factor plus topoisomerase II-targeted drugs in L929 tumor cells. J Natl Cancer Inst 83: 346–350, 1991
  16. Fehsel, K, Kolb-Bachofen, V, Kolb, H: Analysis of TNF alpha induced DNA strand breaks at the single cell level. Am J Pathol 139: 251–254, 1991 | PubMed | ISI | ChemPort |
  17. Gromkowski, SH, Mama, K, Yagi, J, Sen, R, Rath, S: Double stranded RNA and bacterial lipopolysaccharide enhance sensitivity to TNF-alpha-mediated cell death. Int Immunol 2: 903–908, 1990
  18. Fulton, AM, Chong, YC: The role of macrophage-derived TNF alpha in the induction of sublethal tumor cell DNA damage. Carcinogenesis 13: 77–81, 1992
  19. Eisinger, M, Jisootee, J, Hefton, M, Darzynkiewicz, Z, Chiao, JW, de Harven, E: Human epidermal cell cultures: growth and differentiation in the absence dermal components of medium supplements. Proc Natl Acad Sci USA 76: pp 5340–5344, 1979
  20. Boukam, P, Petrussevska, RT, Breitkreutz, D, Hornung, J, Markhan, A, Fusenig, NE: Normal keratinization in a spontaneously immortalized aneuploid human keratinocyte cell line. J Cell Biol 106: 761–771, 1988
  21. Gnvrieli, Y, Sherman, Y, Ben-Sasson, SA: Identification of programmed cell death in situ via specific labeling of nuclear DNA fragmentation. J Cell Biol 119: 493–501, 1992
  22. Gilchrest, BA, Soter, NA, Stoff, JS, Mihm, MC: The human sunburn reaction: histologic and biochemical studies. J Am Acad Dermatol 5: 111–122, 1981
  23. Johnson, HE, Daniels, F Jr: Lysosomes and the reactions of skin to ultraviolet radiation. J Invest Dermatol 53: 85–94, 1969
  24. Brenner, W, Gschnait, H: Decreased DNA repair activity in sunburn cells. Arch Dermatol Res 266: 11–16, 1979 | Article | PubMed | ISI | ChemPort |
  25. Miyachi, Y, Horio, T, Imamura, S: Sunburn cell formation is prevented by scavenging oxygen intermediates. Clin Exp Dermatol 8: 305–310, 1983 | PubMed | ISI | ChemPort |
  26. Danno, K, Horio, T, Takigawa, M, Imamura, S: Role of oxygen intermediates UV-induced epidennal cell injury. J Invets ermatol 83: 166–168, 1984
  27. Youn, JI, Gange, KW, Maytum, D, Parrish, JA: Effect of hypoxia on sunburn cell formation and inflammation induced by ultraviolet radiation. Photodermatology 5: 252–256, 1988 | PubMed | ChemPort |
  28. Larrick, JW, Wright, SC: Cytotoxic mechanism of tumor necrosis factor-alpha. FASEB J 4: 3215–3223, 1990 | PubMed | ISI | ChemPort |
  29. Jarvis, WD, Kolesnick, RN, Fornari, FA, Traylor, RS: Induction of apoptotic DNA damage and cell death by activation of the sphingomyelin pathway. Proc Natl Acad Sci USA 91: 73–77, 1994 | PubMed | ChemPort |
  30. Haimovitz-Friedman, A, Kan, CC, Ehleiter, D, Persaud, RS, McLoughlin, M, Fuks, Z, Kolesnick, RN: Ionizing radiation acts on cellular membranes to generate ceramide and initiate apoptosis. J Exp Med 180: 525–535, 1994 | Article | PubMed | ChemPort |
  31. Lemon, SV, Martin, S, Cotter, TG: Induction of apoptosis (programmed cell death) in tumour cell lines by widely diverging stimuli. Biochem Soc Trans 18: 343–345, 1990
  32. Schwarz, T, Urbanski, A, Luger, TA: Ultraviolet light and epidermal cell-derived cytokines. In: Luger TA, Schwarz T (eds.). Epidermal Growth Factor and Cytokines Marcel Dekker, New York, pp 303–324, 1994
  33. Itoh, N, Yonehara, S, Ishii, A, Yonehara, M, Mizushima, S, Sameshima, M, Hase, A, Seto, Y, Nagata, S: The polypeptide encoded by the cDNA for human cell surface antigen Fas can mediate apoptosis. Cell 66: 233–243, 1991 | Article | PubMed | ISI | ChemPort |
  34. Hockenbery, DM, Zutter, M, Hickey, W, Nahm, M, Korsmeyer, SJ: BCL-2 protein is topographically restricted in tissues characterized by apoptotic cell death. Proc Natl Acad Sci USA 88: 6961–6965, 1991 | PubMed | ChemPort |

Extra navigation

.
ADVERTISEMENT