Skip to main content
SpringerLink
Log in

Increased release of the tumour necrosis factor receptor p75 by immortalized human keratinocytes results from an activated shedding mechanism and is not related to augmented steady-state levels of p75 mRNA

  • Original Paper
  • Published:
Archives of Dermatological Research Aims and scope Submit manuscript

Abstract

The soluble tumour necrosis factor receptor I (sTNFRI, p55) is produced at similar levels by both immortalized (A431, HaCaT, KB) and primary normal human keratinocytes (HNK), whereas the soluble TNFR II (sTNFR II, p75) appears to be specifically released only by immortalized human keratinocytes. The purpose of this study was to investigate whether the increase in p75 secretion by immortalized human keratinocytes is due to an increased shedding of the receptor from the cell membrane, or is related to increased steady-state levels of p75 mRNA. FACS analysis showed that levels of membranous p75 decreased in a time-dependent manner in immortalized cells cultured for 1, 3, 6, 12 and 24 h, while remaining unchanged in HNK throughout. Northern blot analysis showed that after 12 h of culture, when p75 expression was decreased on the cell membrane of all immortalized cells, there was no significant difference in steady state levels of p75 mRNA between immortalized keratinocytes and HNK. Supernatants of immortalized cells, cultured for 24 h contained distinct levels of p75, while levels of p75 in supernatants of HNK were under the detection limit, confirming that the p75 decrease on the cell membrane results from increased p75 shedding from the cell membrane of immortalized cells. In contrast to p75, p55 was continuously expressed on the cell membrane of normal and immortalized keratinocytes without significant variation throughout the entire 24-h culture period and was similarly shed by both cell types. These results suggest that immortalized keratinocytes are specifically activated for shedding of p75 from the cell membrane. Since p75 has a high affinity for TNF, the release of this receptor may imply a direct role in the escape of malignant/transformed keratinocytes from the TNF-mediated immune response.

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. Schuger L, Dixit VM, Carey TE, Varani J (1990) Modulation of squamous carcinoma cell growth, morphology, adhesiveness and extracellular matrix production by interferon-γ and tumour necrosis factor-α. Pathobiology 58:279–286

    Article  PubMed  CAS  Google Scholar 

  2. Erickson SL, Sauvage FJ, Kikly Ch, Carver-Moore K, Pitts-Meek S, Gillet N, Sheehan KCF, Schreiber RD, Goeddel DV, Moore MW (1994) Decreased sensitivity to tumour-necrosis factor but normal T-cell development in TNF receptor-2-deficient mice. Nature 372:560

    Article  PubMed  CAS  Google Scholar 

  3. Brouckaert P, Libert C, Everaerdt B, Fiers W (1992) Selective species specificity of tumour necrosis factor for toxicity in the mouse. Lymphokine Cytokine Res 11:193–196

    PubMed  CAS  Google Scholar 

  4. Aderka D, Wysenbeek A, Engelmann H, Cope, AP, Brennan F, Molad Y, Hornik V, Levo Y, Maini RN, Feldmann M, Wallach D (1993) Correlation between serum levels of soluble tumour necrosis factor receptor and disease activity in systemic lupus erythematosus. Arthritis Rheum 36:1111–1120

    PubMed  CAS  Google Scholar 

  5. Van Zee KJ, Kohno T, Fischer E, Rock CS, Moldawer LL, Lowry SF (1992) Tumour necrosis factor soluble receptors circulate during experimental and clinical inflammation and can protect against excessive tumour necrosis factor α in vitro and in vivo. Proc Natl Acad Sci USA 89:4845

    Article  PubMed  Google Scholar 

  6. Heilig B, Wermann M, Gallati H, Brockhaus M, Berke B, Egen O, Pezzuto A, Hunstein W (1992) Elevated TNF receptor plasma concentrations in patients with rheumatoid arthritis. Clin Invest 70:22–27

    Article  CAS  Google Scholar 

  7. Belldegrun A, Pierce W, Sayah D, deKernion J, Wallach D, Aderka D, Figlin RA (1993) Soluble tumour necrosis factor receptor expression in patients with metastatic renal cell carcinoma treated with interleukin2-based immunotherapy. J Immunother 13:175–180

    PubMed  CAS  Google Scholar 

  8. Gatanaga T, Hwang CD, Kohr W, Cappuccini F, Lucci JA, Jeffes EW, Lentz R, Tomich J, Yamamoto RS, Granger GA (1990) Purification and characterization of an inhibitor (soluble tumour necrosis factor receptor) for tumour necrosis factor and lymphotoxin obtained from the serum ultrafiltrates of human cancer patients. Proc Natl Acad Sci USA 87:8781–8784

    Article  PubMed  CAS  Google Scholar 

  9. Aderka D, Engelmann H, Hornik V, Skornick Y, Levo Y, Wallach D, Kushtai G (1991) Increased serum levels of soluble receptors for tumour necrosis factor in patients. Cancer Res 51:5602–5607

    PubMed  CAS  Google Scholar 

  10. Grosen EA, Granger GA, Gatanaga M, Ininns EK, Hwang C, DiSaia P, Berman M., Manetta A, Emma D, Gatanaga T (1993) Measurement of the soluble membrane receptors for tumour necrosis factor and lymphotoxin in the sera of patients with gynecologic malignancy. Gynecol Oncol 50:68–77

    Article  PubMed  CAS  Google Scholar 

  11. Digel W, Porzsolt F, Schmid M, Herrmann F, Lesslauer W, Brockhaus M (1992) High levels of circulating soluble receptors for tumour necrosis factor in hairy cell leukemia and type B chronic lymphocytic leukemia. J Clin Invest 89:1690–1693

    PubMed  CAS  Google Scholar 

  12. Benelmans MHA, Abramovicz D, Gouma DJ, Goldmann M, Buurmann WA (1994) In vivo T cell activation by anti-CD3 monoclonal antibody induces soluble TNF receptor release in mice. Effects of pentoxifylline, methylprednisolone, anti-TNF, and anti-IFN-γ antibodies. J Immunol 153:499

    Google Scholar 

  13. Upton C, Macen JL, Schreiber M, McFadden G (1991) Myxoma virus expresses a secreted protein with homology to the tumour necrosis factor receptor gene family that contributes to viral virulence. Virology 184:370–382

    Article  PubMed  CAS  Google Scholar 

  14. Neuner P, Klosner G, Pourmojib M, Trautinger F, Knobler R (1994) Selective release of tumour necrosis factor binding protein II by malignant human epidermal cells reveals protection from tumour necrosis factor alpha mediated cytotoxicity. Cancer Res 54:6001–6005

    PubMed  CAS  Google Scholar 

  15. Pillai S, Bikle DD, Eessalu TE, Aggarwal BB, Elias PM (1989) Binding and biological effects of tumour necrosis factor alpha on cultured human neonatal foreskin keratinocytes. J Clin Invest 83:816–821

    PubMed  CAS  Google Scholar 

  16. Old LJ (1987) Tumour necrosis factor. Polypeptide mediator network. Nature 326:330

    Article  PubMed  CAS  Google Scholar 

  17. Trefzer U, Brockhaus M, Lotscher H, Parlow F, Budnik A, Grewe M, Christoph H, Kapp A, Schöpf E, Luger TA, Krutmann J (1993) The 55-kD tumour necrosis factor receptor on human keratinocytes is regulated by tumour necrosis factor-alpha and by ultraviolet B radiation. J Clin Invest 92:462–470

    PubMed  CAS  Google Scholar 

  18. Kirnbauer R, Köck A, Schwarz T, Urbanski A, Krutmann J, Borth W, Damm D, Shipley 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 epidermal cells and epidermoid carcinoma cell lines. J Immunol 142:1922–1928

    PubMed  CAS  Google Scholar 

  19. Trefzer U, Brockhaus M, Loetscher H, Parlow F, Kapp A, Schöpf E, Krutmann J (1991) 55-kD tumour necrosis factor receptor is expressed by human keratinocytes and plays a pivotal role in regulation of human keratinocyte ICAM-1 expression. J Invest Dermatol 97:911–916

    Article  PubMed  CAS  Google Scholar 

  20. Björnberg F, Lantz N, Olsson I, Gullberg U (1994) Mechanisms involved in the processing of the p55 and the p75 tumour necrosis factor (TNF) receptors to soluble receptor forms. Lymphokine Cytokine Res 13:203–211

    PubMed  Google Scholar 

  21. Nophar Y, Kemper O, Brakebusch C, Englemann H, Zwang R, Aderka D, Holtmann H, Wallach D (1990) Soluble forms of tumour necrosis factor receptors (TNF-Rs). The cDNA for the type I TNF-R, cloned using amino acid sequence data of its soluble form, encodes both the cell surface and a soluble form of the receptor. EMBO J 9:3269–3278

    PubMed  CAS  Google Scholar 

  22. Heller RA, Song K, Onasch MA, Fischer WH, Chang D, Ringold GM (1990) Complementery DNA cloning of a receptor for tumour necrosis factor and demonstration of a shed form of the receptor. Proc Natl Acad Sci USA 87:6151–6155

    Article  PubMed  CAS  Google Scholar 

  23. Lantz N, Gullberg U, Nilsson E, Olsson I (1990) Characterization in vitro of a human tumour necrosis factor-binding protein. A soluble form of a tumour necrosis factor receptor. J Clin Invest 86:1396–1402

    Article  PubMed  CAS  Google Scholar 

  24. Kohno T, Brewer MT, Baker SL, Schwartz PE, King MW, Hale KK, Squires CH, Thompson RC, Vannice JL (1990) A second tumour necrosis factor receptor gene product can shed a naturally occurring tumour necrosis factor inhibitor. Proc Natl Acad Sci USA 87:8331–8335

    Article  PubMed  CAS  Google Scholar 

  25. Crowe PK, Van Arsdale TL, Goodwin RG, Ware CF (1993) Specific induction of 80-kDa tumour necrosis factor receptor shedding in T lymphocytes involves the cytoplasmic domain and phosphorylation. J Immunol 151:6882–6890

    PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Dermatology, Division of Special and Environmental Dermatology, University of Vienna, A-1090, Vienna, Austria

    Peter Neuner, Mojgan Pourmojib, Gabriele Klosner, Franz Trautinger & Robert Knobler

Authors
  1. Peter Neuner
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Mojgan Pourmojib
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Gabriele Klosner
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Franz Trautinger
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Robert Knobler
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

This work was supported by a grant from the ‘Medizinisch-Wissenschaftlicher Fonds des Bürgermeisters der Bundeshauptstadt Wien’ and the ‘Hochschuljubiläumsstiftung der Stadt Wien’

Rights and permissions

Reprints and permissions

About this article

Cite this article

Neuner, P., Pourmojib, M., Klosner, G. et al. Increased release of the tumour necrosis factor receptor p75 by immortalized human keratinocytes results from an activated shedding mechanism and is not related to augmented steady-state levels of p75 mRNA. Arch Dermatol Res 288, 691–696 (1996). https://doi.org/10.1007/BF02505279

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02505279

Key words

Search

Navigation