Archives of Dermatological Research

, Volume 286, Issue 2, pp 73–76 | Cite as

Influence of various cytokines on the interleukin-2-dependent lysis of melanoma cells in vitro

  • E. S. Schultz
  • R. Dummer
  • J. C. Becker
  • D. Zillikens
  • G. Burg
Original Contributions


To obtain information about useful combinations of various cytokines in melanoma therapy, we studied the influence of interleukin-2 (IL-2) in combination with interferon-alpha (IFN-alpha), IFN-gamma and tumour necrosis factor-alpha (TNF-alpha) on the lytic activity of IL-2-stimulated cells in vitro. Peripheral mononuclear cells (PMC) were incubated for 4 days with various combinations of cytokines and used as effector cells. Two different melanoma cell lines (M19 and M26) were used as target cells. The lytic activity of stimulated PMC was determined using a modified hexosaminidase assay. IL-2 was mainly responsible for the lytic activity of the effector cells in a dose-dependent manner. IFN-alpha, IFN-gamma and TNF-alpha did not enhance lytic activity with an optimal IL-2 dose (50 IU/ml IL-2). Using a suboptimal IL-2 dose (5 IU/ml), they increased cytotoxicity. The specific lysis of M19 cells was significantly increased by pretreatment of the cells with 5 IU/ml IFN-alpha together with 50 IU/ml TNF-alpha (t-test, P≤0.001), while the specific lysis of M26 cells was increased by pretreatment with 5 IU/ml IFN-gamma. We conclude that the lysis of melanoma cells by cytotoxic cells in vitro can be enhanced by various cytokines. The optimal cytokine combination differed for the two melanoma cell lines tested.

Key words

Melanoma Interleukin-2 Interferon Tumour necrosis factor Cytokines 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Atzpodien J, Körfer A, Franks CR, Polidowa H, Kirchner H (1990) Home therapy with recombinant interleukin-2 and interferon-alpha-2b in advanced human malignancies. Lancet 335: 1509–1512Google Scholar
  2. 2.
    Bergmann L (1989) Malignant melanoma — prognosis and actual treatment strategies with chemotherapy and biological response modifiers. Eur J Cancer Clin Oncol 25 [Suppl 3]: S31-S36Google Scholar
  3. 3.
    Bergmann L, Weidmann E, Bungert B, Hechler P, Mitrou PS (1990) Influence of various cytokines on the induction of lymphokine-activated killer cells. Nat Immun Cell Growth Regul 9: 265–273Google Scholar
  4. 4.
    Carswell EA, Old LJ, Kassel RL (1975) An endotoxin induced serum factor that causes necrosis of tumors. Proc Natl Acad Sci USA 72: 3666–3670Google Scholar
  5. 5.
    Chouaib S (1991) Tumor necrosis factor alpha: costimulator for cytotoxic cell differentiation. Nouv Rev Fr Hematol 33 (6): 471–475Google Scholar
  6. 6.
    De Fries RU, Golub SH (1988) Characteristics and mechanism of IFN-gamma induced protection of human tumor cells from lysis by lymphokine-activated killer cells. J Immunol 140: 3686–3693Google Scholar
  7. 7.
    Garbe C, Krasagakis K, Zouboulis CC, Schroder K, Krüger S, Stadler R, Orfanos CE (1990) Antitumor activities of interferon alpha, beta, and gamma and their combinations on human melanoma cells in vitro: change of proliferation, melanin synthesis and immunophenotype. J Invest Dermatol 95 (6 Suppl): 231–237Google Scholar
  8. 8.
    Hirsh M, Lipton A, Harvey H, Givant E, Hopper K, Jones G, Zeffren J, Levitt D (1990) Phase I study of interleukin-2 and interferon alfa-2a as outpatient therapy for patients with advanced malignancy. J Clin Oncol 8 (10): 1657–1663Google Scholar
  9. 9.
    Kaufmann Y, Davidson J, Levanon M, Icekson I, Revel M, Ramot B (1991) Lymphokine-activated killer (LAK) cells: interferon-gamma synergizes with interleukin-2 to induce LAK cytotoxicity in homogeneous leukemic preparations. Clin Immunol Immunopathol 58 (2): 278–288Google Scholar
  10. 10.
    Krüger S, Garbe C, Orfanos CE (1990) A new fluorometric microassay for the determination of melanoma cell (MC) susceptibility to LAK cell lysis (abstract). J Invest Dermatol 95 (4): 476Google Scholar
  11. 11.
    Lafraniere R, Rosenberg SA (1985) Adoptive immunotherapy of murine hepatic metastases with lymphokine activated killer (LAK) cells and recombinant interleukin-2 (rIL-2) can mediate the regression of both immunogenic and nonimmunogenic sarcomas and an adenocarcinoma. J Immunol 135: 4273–4280Google Scholar
  12. 12.
    Landgren V (1984) Measurement of cell numbers by means of the endogenous enzyme hexosaminidase. Applications to detection of lymphokines and cell surface antigens. J Immunol Meth 67: 379–388Google Scholar
  13. 13.
    Maghazachi AA (1990) Differential effects of various cytokines on the generation of rat LAK cells from their purified precursors. Immunology 70 (4): 465–472Google Scholar
  14. 14.
    Matossian-Rogers A, Browne C, Turkish M, O'Byrne P, Festenstein H (1989) Tumour necrosis factor-alpha enhances the cytolytic and cytostatic capacity of interleukin-2 activated killer cells. Br J Cancer 59: 573–577Google Scholar
  15. 15.
    Muss HB (1988) Interferon therapy of metastatic renal cell carcinoma. Semin Surg Oncol 4: 199–203Google Scholar
  16. 16.
    Négrier S, Philip T, Stoter G, et al. (1989) Interleukin-2 with or without LAK cells in metastatic renal cell carcinoma: a report of European multicenter study. Eur J Cancer Clin Oncol 25 [Suppl 3]: S21-S28Google Scholar
  17. 17.
    Parkinson DR, Abrams JS, Wiernik PH, et al (1990) Interleukin-2 therapy in patients with metastatic malignant melanoma: A phase II study. J Clin Oncol 8 (10): 1650–1656Google Scholar
  18. 18.
    Rosenberg SA (1988) Cancer therapy with interleukin-2: immunological manipulations mediate the regression of cancer in humans. J Clin Oncol 6: 402–405Google Scholar
  19. 19.
    Rosenberg SA, Lotze MT, Yang JC, et al (1989) Experience with the use of high-dose interleukin-2 in the treatment of 652 cancer patients. Ann Surg 210: 474–485Google Scholar
  20. 20.
    Rosenberg SA, Lotze MT, Yang JC, Linehan WM, Seipp C, Calabro S, Karp SE, Sherry RM, Steinberg S, White DE (1989) Combination therapy with interleukin-2 and alpha-interferon for the treatment of patients with advanced cancer. J Clin Oncol 7 (12): 1863–1874Google Scholar
  21. 21.
    Sone S, Utsugi T, Nii A, Ogura T (1988) Differential effects of recombinant interferons alpha, beta, and gamma on induction of human lymphokine (IL-2)-activated killer activity. J. Nat Cancer Inst 80 (6): 425–431Google Scholar
  22. 22.
    Versteeg R (1992) NK cells and T cells: mirror images? Immunol Today 13 (7): 244–247Google Scholar

Copyright information

© Springer-Verlag 1994

Authors and Affiliations

  • E. S. Schultz
    • 1
  • R. Dummer
    • 1
  • J. C. Becker
    • 2
  • D. Zillikens
    • 2
  • G. Burg
    • 1
  1. 1.Department of DermatologyUniversity of Zurich Medical SchoolZurichSwitzerland
  2. 2.Department of DermatologyUniversity of Würzburg Medical SchoolWürzburgGermany

Personalised recommendations