Cancer Immunology, Immunotherapy

, Volume 60, Issue 1, pp 123–131 | Cite as

Resistance to the proapoptotic effects of interferon-gamma on melanoma cells used in patient-specific dendritic cell immunotherapy is associated with improved overall survival

  • A. N. Cornforth
  • A. W. Fowler
  • D. J. Carbonell
  • R. O. Dillman
Original Article


The use of whole cell tumor vaccines and various means of loading antigen onto dendritic cells have been under investigation for over a decade. Induction of apoptosis and the exposure of immune-stimulating proteins are thought to be beneficial for the use in immunotherapy protocols, but conclusive evidence in the clinical setting has been lacking. Incubation of melanoma cell lines with interferon-gamma (IFN-γ) increased phosphatidylserine and calreticulin exposure, but not in the IFN-γ-resistant cell line Lu-1205. Short-term autologous melanoma cell lines used for loading dendritic cells for immunotherapy showed differential response to the pro-apoptotic effects of IFN-γ. These IFN-γ-treated tumor cells (TCs) were irradiated and used for loading antigen for dendritic cell therapy. A log-rank comparison of survival for patients whose TCs were found to be either sensitive (upregulated phosphatidylserine and calreticulin) or insensitive to IFN-γ revealed a strongly significant correlation to progression-free (p = 0.003) and overall survival (p = 0.002) favorably in those patients whose cell lines were resistant to the proapoptotic effect of IFN-γ. These results suggest that the use of IFN-γ in anti-melanoma dendritic cell-based immunotherapy may only be beneficial when the cells do not undergo apoptosis in response to IFN-γ and support the contention that the use of some apoptotic cells in vaccines may be detrimental.


Dendritic cells Immunotherapy Interferon-gamma Apoptosis Melanoma 



We would like to acknowledge Sarah Tillman and Andrea Beatty for their assistance in generating the tumor and dendritic cell lines and Amy Mattes for assistance in the cell death assays. Funding for this work was provided by the Hoag Hospital Foundation.


  1. 1.
    Palucka AK, Ueno H, Fay JW, Banchereau J (2007) Taming cancer by inducing immunity via dendritic cells. Immunol Rev 220:129–150CrossRefPubMedGoogle Scholar
  2. 2.
    Tuyaerts S, Aerts JL, Corthals J, Neyns B, Heirman C, Breckpot K et al (2007) Current approaches in dendritic cell generation and future implications for cancer immunotherapy. Cancer Immunol Immunother 56:1513–1537CrossRefPubMedGoogle Scholar
  3. 3.
    Cranmer LD, Trevor KT, Hersh EM (2004) Clinical applications of dendritic cell vaccination in the treatment of cancer. Cancer Immunol Immunother 53:275–306CrossRefPubMedGoogle Scholar
  4. 4.
    Dillman RO, Selvan SR, Schiltz PM (2006) Patient-specific dendritic-cell vaccines for metastatic melanoma. N Engl J Med 355:1179–1181CrossRefPubMedGoogle Scholar
  5. 5.
    Dillman RO, Nayak SK, Barth NM, DeLeon C, Schwartzberg LS, Spitler LE et al (1998) Clinical experience with autologous tumor cell lines for patient-specific vaccine therapy in metastatic melanoma. Cancer Biother Radiopharm 13:165–176CrossRefPubMedGoogle Scholar
  6. 6.
    Meyskens FL Jr, Kopecky KJ, Taylor CW, Noyes RD, Tuthill RJ, Hersh EM et al (1995) Randomized trial of adjuvant human interferon gamma versus observation in high-risk cutaneous melanoma: a Southwest Oncology Group Study. J Natl Cancer Inst 87:1710–1713CrossRefPubMedGoogle Scholar
  7. 7.
    Lotem M, Machlenkin A, Hamburger T, Nissan A, Kadouri L, Frankenburg S et al (2009) Autologous melanoma vaccine induces antitumor and self-reactive immune responses that affect patient survival and depend on MHC class II expression on vaccine cells. Clin Cancer Res 15:4968–4977CrossRefPubMedGoogle Scholar
  8. 8.
    Fujisawa Y, Nabekura T, Nakao T, Nakamura Y, Takahashi T, Kawachi Y et al (2009) The induction of tumor-specific CD4+ T cells via major histocompatibility complex class II is required to gain optimal anti-tumor immunity against B16 melanoma cell line in tumor immunotherapy using dendritic cells. Exp Dermatol 18:396–403CrossRefPubMedGoogle Scholar
  9. 9.
    Zoller M, Strubel A, Hammerling G, Andrighetto G, Raz A, Ben-Ze’ev A (1988) Interferon-gamma treatment of B16 melanoma cells: opposing effects for non-adaptive and adaptive immune defense and its reflection by metastatic spread. Int J Cancer 41:256–266CrossRefPubMedGoogle Scholar
  10. 10.
    Ugurel S, Seiter S, Rappl G, Stark A, Tilgen W, Reinhold U (1999) Heterogenous susceptibility to CD95-induced apoptosis in melanoma cells correlates with bcl-2 and bcl-x expression and is sensitive to modulation by interferon-gamma. Int J Cancer 82:727–736CrossRefPubMedGoogle Scholar
  11. 11.
    Ullrich E, Bonmort M, Mignot G, Kroemer G, Zitvogel L (2008) Tumor stress, cell death and the ensuing immune response. Cell Death Differ 15:21–28CrossRefPubMedGoogle Scholar
  12. 12.
    Sporri R, Reis e Sousa C (2005) Inflammatory mediators are insufficient for full dendritic cell activation and promote expansion of CD4+ T cell populations lacking helper function. Nat Immunol 6:163–170CrossRefPubMedGoogle Scholar
  13. 13.
    Obeid M, Panaretakis T, Joza N, Tufi R, Tesniere A, van Endert P et al (2007) Calreticulin exposure is required for the immunogenicity of gamma-irradiation and UVC light-induced apoptosis. Cell Death Differ 14:1848–1850CrossRefPubMedGoogle Scholar
  14. 14.
    Fry TJ, Shand JL, Milliron M, Tasian SK, Mackall CL (2009) Antigen loading of DCs with irradiated apoptotic tumor cells induces improved anti-tumor immunity compared to other approaches. Cancer Immunol Immunother 58:1257–1264CrossRefPubMedGoogle Scholar
  15. 15.
    Obeid M, Tesniere A, Ghiringhelli F, Fimia GM, Apetoh L, Perfettini JL et al (2007) Calreticulin exposure dictates the immunogenicity of cancer cell death. Nat Med 13:54–61CrossRefPubMedGoogle Scholar
  16. 16.
    Del Cid N, Jeffery E, Rizvi SM, Stamper E, Peters LR, Brown WC et al (2010) Modes of calreticulin recruitment to the major histocompatibility complex class I assembly pathway. J Biol Chem 285:4520–4535CrossRefPubMedGoogle Scholar
  17. 17.
    Su Q, Wang S, Baltzis D, Qu LK, Raven JF, Li S et al (2007) Interferons induce tyrosine phosphorylation of the eIF2alpha kinase PKR through activation of Jak1 and Tyk2. EMBO Rep 8:265–270CrossRefPubMedGoogle Scholar
  18. 18.
    Dillman RO, Selvan SR, Schiltz PM, McClay EF, Barth NM, DePriest C et al (2009) Phase II trial of dendritic cells loaded with antigens from self-renewing, proliferating autologous tumor cells as patient-specific antitumor vaccines in patients with metastatic melanoma: final report. Cancer Biother Radiopharm 24:311–319CrossRefPubMedGoogle Scholar
  19. 19.
    Choi D, Perrin M, Hoffmann S, Chang AE, Ratanatharathorn V, Uberti J et al (1998) Dendritic cell-based vaccines in the setting of peripheral blood stem cell transplantation: CD34+ cell-depleted mobilized peripheral blood can serve as a source of potent dendritic cells. Clin Cancer Res 4:2709–2716PubMedGoogle Scholar
  20. 20.
    Luft T, Pang KC, Thomas E, Bradley CJ, Savoia H, Trapani J et al (1998) A serum-free culture model for studying the differentiation of human dendritic cells from adult CD34+ progenitor cells. Exp Hematol 26:489–500PubMedGoogle Scholar
  21. 21.
    Dillman RO, Nayak SK, Beutel L (1993) Establishing in vitro cultures of autologous tumor cells for use in active specific immunotherapy. J Immunother Emphasis Tumor Immunol 14:65–69PubMedGoogle Scholar
  22. 22.
    Dillman RO, Beutel LD, Cornforth AN, Nayak SK (2000) Short-term tumor cell lines from renal cell carcinoma for use as autologous tumor cell vaccines in the treatment of kidney cancer. Cancer Biother Radiopharm 15:161–168CrossRefPubMedGoogle Scholar
  23. 23.
    Selvan SR, Dillman RO, Fowler AW, Carbonell DJ, Ravindranath MH (2007) Monitoring response to treatment in melanoma patients: potential of a serum glycomic marker. Int J Cancer 122(6):1374–1383CrossRefGoogle Scholar
  24. 24.
    Selvan SR, Carbonell DJ, Fowler AW, Beatty AR, Ravindranath MH, Dillman RO (2010) Establishment of stable cell lines for personalized melanoma cell vaccine. Melanoma Res 20(4)280–292Google Scholar
  25. 25.
    Fojtova M, Boudny V, Kovarik A, Lauerova L, Adamkova L, Souckova K et al (2007) Development of IFN-gamma resistance is associated with attenuation of SOCS genes induction and constitutive expression of SOCS 3 in melanoma cells. Br J Cancer 97:231–237CrossRefPubMedGoogle Scholar
  26. 26.
    Panaretakis T, Kepp O, Brockmeier U, Tesniere A, Bjorklund AC, Chapman DC et al (2009) Mechanisms of pre-apoptotic calreticulin exposure in immunogenic cell death. EMBO J 28:578–590CrossRefPubMedGoogle Scholar
  27. 27.
    Adema GJ, de Vries IJ, Punt CJ, Figdor CG (2005) Migration of dendritic cell based cancer vaccines: in vivo veritas? Curr Opin Immunol 17:170–174CrossRefPubMedGoogle Scholar
  28. 28.
    Kepp O, Tesniere A, Schlemmer F, Michaud M, Senovilla L, Zitvogel L et al (2009) Immunogenic cell death modalities and their impact on cancer treatment. Apoptosis 14:364–375CrossRefPubMedGoogle Scholar
  29. 29.
    Martins I, Tesniere A, Kepp O, Michaud M, Schlemmer F, Senovilla L et al (2009) Chemotherapy induces ATP release from tumor cells. Cell Cycle 8:3723–3728CrossRefPubMedGoogle Scholar
  30. 30.
    Apetoh L, Ghiringhelli F, Tesniere A, Criollo A, Ortiz C, Lidereau R et al (2007) The interaction between HMGB1 and TLR4 dictates the outcome of anticancer chemotherapy and radiotherapy. Immunol Rev 220:47–59CrossRefPubMedGoogle Scholar
  31. 31.
    Blank C, Gajewski TF, Mackensen A (2005) Interaction of PD-L1 on tumor cells with PD-1 on tumor-specific T cells as a mechanism of immune evasion: implications for tumor immunotherapy. Cancer Immunol Immunother 54:307–314CrossRefPubMedGoogle Scholar
  32. 32.
    Dong H, Strome SE, Salomao DR, Tamura H, Hirano F, Flies DB et al (2002) Tumor-associated B7-H1 promotes T-cell apoptosis: a potential mechanism of immune evasion. Nat Med 8:793–800PubMedGoogle Scholar
  33. 33.
    Hemmi H, Akira S (2005) TLR signalling and the function of dendritic cells. Chem Immunol Allergy 86:120–135CrossRefPubMedGoogle Scholar
  34. 34.
    Marin-Gallen S, Clemente-Casares X, Planas R, Pujol-Autonell I, Carrascal J, Carrillo J et al (2010) Dendritic cells pulsed with antigen-specific apoptotic bodies prevent experimental type 1 diabetes. Clin Exp Immunol 160:207–214CrossRefPubMedGoogle Scholar
  35. 35.
    Ferguson TA, Kazama H (2005) Signals from dying cells: tolerance induction by the dendritic cell. Immunol Res 32:99–108CrossRefPubMedGoogle Scholar
  36. 36.
    Kamei T, Inui M, Nakamura S, Okumura K, Goto A, Tagawa T (2003) Interferon-gamma and anti-Fas antibody-induced apoptosis in human melanoma cell lines and its relationship to bcl-2 cleavage and bak expression. Melanoma Res 13:153–159CrossRefPubMedGoogle Scholar
  37. 37.
    Tesniere A, Panaretakis T, Kepp O, Apetoh L, Ghiringhelli F, Zitvogel L et al (2008) Molecular characteristics of immunogenic cancer cell death. Cell Death Differ 15:3–12CrossRefPubMedGoogle Scholar
  38. 38.
    Perez CA, Fu A, Onishko H, Hallahan DE, Geng L (2009) Radiation induces an antitumour immune response to mouse melanoma. Int J Radiat Biol 85:1126–1136CrossRefPubMedGoogle Scholar
  39. 39.
    Dillman RO, DePriest C, DeLeon C, Barth NM, Schwartzberg LS, Beutel LD et al (2007) Patient-specific vaccines derived from autologous tumor cell lines as active specific immunotherapy: results of exploratory phase I/II trials in patients with metastatic melanoma. Cancer Biother Radiopharm 22:309–321CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag 2010

Authors and Affiliations

  • A. N. Cornforth
    • 1
  • A. W. Fowler
    • 1
  • D. J. Carbonell
    • 1
  • R. O. Dillman
    • 1
  1. 1.Cell Biology Laboratory, Hoag Cancer CenterNewport BeachUSA

Personalised recommendations