Advertisement

Cancer Immunology, Immunotherapy

, Volume 64, Issue 7, pp 861–872 | Cite as

Development of a potent melanoma vaccine capable of stimulating CD8+ T-cells independently of dendritic cells in a mouse model

  • Katie L. PowellEmail author
  • Alexandre S. Stephens
  • Stephen J. Ralph
Original Article

Abstract

At present, there are no vaccines approved for the prevention or treatment of malignant melanoma, despite the amount of time and resources that has been invested. In this study, we aimed to develop a self-contained vaccine capable of directly stimulating anticancer CD8+ T-cell immune responses. To achieve this, three whole-cell melanoma vaccines were developed expressing 4-1BBL or B7.1 T-cell co-stimulatory molecules individually or in combination. The ability of engineered vaccine cell lines to stimulate potent anticancer immune responses in C57BL/6 mice was assessed. Mice vaccinated with cells overexpressing both 4-1BBL and B7.1 (B16-F10-4-1BBL-B7.1-IFNγ/β anticancer vaccine) displayed the greatest increases in CD8+ T-cell populations (1.9-fold increase versus control within spleens), which were efficiently activated following antigenic stimulation, resulting in a 10.7-fold increase in cancer cell cytotoxicity relative to control. The enhanced immune responses in B16-F10-4-1BBL-B7.1-IFNγ/β-vaccinated mice translated into highly efficient rejection of live tumour burdens and conferred long-term protection against repeated tumour challenges, which were likely due to enhanced effector memory T-cell populations. Similar results were observed when dendritic cell (DC)-deficient LTα−/− mice were treated with the B16-F10-4-1BBL-B7.1-IFNγ/β anticancer vaccine, suggesting that the vaccine can directly stimulate CD8+ T-cell responses in the context of severely reduced DCs. This study shows that the B16-F10-4-1BBL-B7.1-IFNγ/β anticancer vaccine acted as a highly effective antigen-presenting cell and is likely to be able to directly stimulate CD8+ T-cells, without requiring co-stimulatory signals from either CD4+ T-cells or DCs, and warrants translation of this technology into the clinical setting.

Keywords

Vaccine Melanoma Immune response Cytotoxic T-cells Effector memory CD8+ T-cells 

Abbreviations

APC

Antigen-presenting cell

CTL

Cytotoxic T lymphocyte

DC

Dendritic cell

IFN

Interferon

MLC

Mixed lymphocyte culture

TCM

Central memory CD8+ T-cell

TEM

Effector memory CD8+ T-cell

Notes

Acknowledgments

The authors would like to acknowledge Dr. Brendan Hill and Nigel Middlebrooke at Premion Cancer Care, Southport, for assisting with the use of the irradiator. This study was supported by Genvax Pty. Ltd. Katie L. Powell was supported by an Australian Postgraduate Award.

Conflict of interest

S.J. Ralph is the Director of Genvax Pty. Ltd., and this study was supported by funding from Genvax Pty. Ltd. S.J. Ralph is a consultant and inventor on patents related to the commercialisation of cancer vaccines including some technology described herein. S.J. Ralph and K.L. Powell have filed a patent relating to the material presented. A.S. Stephens has no conflict of interest to declare.

Supplementary material

262_2015_1695_MOESM1_ESM.pdf (200 kb)
Supplementary material 1 (PDF 200 kb)

References

  1. 1.
    Trotter SC, Sroa N, Winkelmann RR, Olencki T, Bechtel M (2013) A global review of melanoma follow-up guidelines. J Clin Aesthet Dermatol 6:18–26PubMedCentralPubMedGoogle Scholar
  2. 2.
    Bray F, Jemal A, Grey N, Ferlay J, Forman D (2012) Global cancer transitions according to the Human Development Index (2008–2030): a population-based study. Lancet Oncol 13:790–801PubMedCrossRefGoogle Scholar
  3. 3.
    Azoury SC, Lange JR (2014) Epidemiology, risk factors, prevention, and early detection of melanoma. Surg Clin North Am 94:945–962, viiPubMedCrossRefGoogle Scholar
  4. 4.
    Lee C, Collichio F, Ollila D, Moschos S (2013) Historical review of melanoma treatment and outcomes. Clin Dermatol 31:141–147PubMedCrossRefGoogle Scholar
  5. 5.
    Fang L, Lonsdorf AS, Hwang ST (2008) Immunotherapy for advanced melanoma. J Invest Dermatol 128:2596–2605PubMedCrossRefGoogle Scholar
  6. 6.
    Berd D, Maguire HC Jr, Schuchter LM, Hamilton R, Hauck WW, Sato T, Mastrangelo MJ (1997) Autologous hapten-modified melanoma vaccine as postsurgical adjuvant treatment after resection of nodal metastases. J Clin Oncol 15:2359–2370PubMedGoogle Scholar
  7. 7.
    Soiffer R, Hodi FS, Haluska F et al (2003) Vaccination with irradiated, autologous melanoma cells engineered to secrete granulocyte-macrophage colony-stimulating factor by adenoviral-mediated gene transfer augments antitumor immunity in patients with metastatic melanoma. J Clin Oncol 21:3343–3350PubMedCrossRefGoogle Scholar
  8. 8.
    O’Rourke MG, Johnson M, Lanagan C et al (2003) Durable complete clinical responses in a phase I/II trial using an autologous melanoma cell/dendritic cell vaccine. Cancer Immunol Immunother 52:387–395PubMedGoogle Scholar
  9. 9.
    de Rosa F, Ridolfi L, Ridolfi R et al (2014) Vaccination with autologous dendritic cells loaded with autologous tumor lysate or homogenate combined with immunomodulating radiotherapy and/or preleukapheresis IFN-alpha in patients with metastatic melanoma: a randomised “proof-of-principle” phase II study. J Transl Med 12:209PubMedCentralPubMedCrossRefGoogle Scholar
  10. 10.
    Jaffee EM, Hruban RH, Biedrzycki B et al (2001) Novel allogeneic granulocyte-macrophage colony-stimulating factor-secreting tumor vaccine for pancreatic cancer: a phase I trial of safety and immune activation. J Clin Oncol 19:145–156PubMedGoogle Scholar
  11. 11.
    Sondak VK, Sabel MS, Mule JJ (2006) Allogeneic and autologous melanoma vaccines: where have we been and where are we going? Clin Cancer Res 12:2337s–2341sPubMedCrossRefGoogle Scholar
  12. 12.
    Ralph SJ (2007) An update on malignant melanoma vaccine research: insights into mechanisms for improving the design and potency of melanoma therapeutic vaccines. Am J Clin Dermatol 8:123–141PubMedCrossRefGoogle Scholar
  13. 13.
    Dezfouli S, Hatzinisiriou I, Ralph SJ (2003) Enhancing CTL responses to melanoma cell vaccines in vivo: synergistic increases obtained using IFNgamma primed and IFNbeta treated B7-1+ B16-F10 melanoma cells. Immunol Cell Biol 81:459–471PubMedCrossRefGoogle Scholar
  14. 14.
    Hsueh EC, Morton DL (2003) Antigen-based immunotherapy of melanoma: canvaxin therapeutic polyvalent cancer vaccine. Semin Cancer Biol 13:401–407PubMedCrossRefGoogle Scholar
  15. 15.
    Faries MB, Hsueh EC, Ye X, Hoban M, Morton DL (2009) Effect of granulocyte/macrophage colony-stimulating factor on vaccination with an allogeneic whole-cell melanoma vaccine. Clin Cancer Res 15:7029–7035PubMedCentralPubMedCrossRefGoogle Scholar
  16. 16.
    Lotem M, Kadouri L, Merims S et al (2011) HLA-B35 correlates with a favorable outcome following adjuvant administration of an HLA-matched allogeneic melanoma vaccine. Tissue Antigens 78:203–207PubMedCrossRefGoogle Scholar
  17. 17.
    Mogi S, Sakurai J, Kohsaka T, Enomoto S, Yagita H, Okumura K, Azuma M (2000) Tumour rejection by gene transfer of 4-1BB ligand into a CD80(+) murine squamous cell carcinoma and the requirements of co-stimulatory molecules on tumour and host cells. Immunology 101:541–547PubMedCentralPubMedCrossRefGoogle Scholar
  18. 18.
    DeBenedette MA, Wen T, Bachmann MF, Ohashi PS, Barber BH, Stocking KL, Peschon JJ, Watts TH (1999) Analysis of 4-1BB ligand (4-1BBL)-deficient mice and of mice lacking both 4-1BBL and CD28 reveals a role for 4-1BBL in skin allograft rejection and in the cytotoxic T cell response to influenza virus. J Immunol 163:4833–4841PubMedGoogle Scholar
  19. 19.
    Stannard KA, Collins PM, Ito K et al (2010) Galectin inhibitory disaccharides promote tumour immunity in a breast cancer model. Cancer Lett 299:95–110PubMedCrossRefGoogle Scholar
  20. 20.
    Townsend SE, Allison JP (1993) Tumor rejection after direct costimulation of CD8+ T cells by B7-transfected melanoma cells. Science 259:368–370PubMedCrossRefGoogle Scholar
  21. 21.
    Cosgrove D, Gray D, Dierich A, Kaufman J, Lemeur M, Benoist C, Mathis D (1991) Mice lacking MHC class II molecules. Cell 66:1051–1066PubMedCrossRefGoogle Scholar
  22. 22.
    De Togni P, Goellner J, Ruddle NH et al (1994) Abnormal development of peripheral lymphoid organs in mice deficient in lymphotoxin. Science 264:703–707PubMedCrossRefGoogle Scholar
  23. 23.
    Wu Q, Wang Y, Wang J, Hedgeman EO, Browning JL, Fu YX (1999) The requirement of membrane lymphotoxin for the presence of dendritic cells in lymphoid tissues. J Exp Med 190:629–638PubMedCentralPubMedCrossRefGoogle Scholar
  24. 24.
    Steinman RM (1991) The dendritic cell system and its role in immunogenicity. Annu Rev Immunol 9:271–296PubMedCrossRefGoogle Scholar
  25. 25.
    Ardavin C, del Hoyo GM, Martin P, Anjuere F, Arias CF, Marin AR, Ruiz S, Parrillas V, Hernandez H (2001) Origin and differentiation of dendritic cells. Trends Immunol 22:691–700PubMedCrossRefGoogle Scholar
  26. 26.
    Gabrilovich DI, Nadaf S, Corak J, Berzofsky JA, Carbone DP (1996) Dendritic cells in antitumor immune responses. II. Dendritic cells grown from bone marrow precursors, but not mature DC from tumor-bearing mice, are effective antigen carriers in the therapy of established tumors. Cell Immunol 170:111–119PubMedCrossRefGoogle Scholar
  27. 27.
    Almand B, Resser JR, Lindman B, Nadaf S, Clark JI, Kwon ED, Carbone DP, Gabrilovich DI (2000) Clinical significance of defective dendritic cell differentiation in cancer. Clin Cancer Res 6:1755–1766PubMedGoogle Scholar
  28. 28.
    Tas MP, Simons PJ, Balm FJ, Drexhage HA (1993) Depressed monocyte polarization and clustering of dendritic cells in patients with head and neck cancer: in vitro restoration of this immunosuppression by thymic hormones. Cancer Immunol Immunother 36:108–114PubMedCrossRefGoogle Scholar
  29. 29.
    Gabrilovich DI, Corak J, Ciernik IF, Kavanaugh D, Carbone DP (1997) Decreased antigen presentation by dendritic cells in patients with breast cancer. Clin Cancer Res 3:483–490PubMedGoogle Scholar
  30. 30.
    O’Rourke MG, Johnson MK, Lanagan CM et al (2007) Dendritic cell immunotherapy for stage IV melanoma. Melanoma Res 17:316–322PubMedCrossRefGoogle Scholar
  31. 31.
    Trepiakas R, Berntsen A, Hadrup SR et al (2010) Vaccination with autologous dendritic cells pulsed with multiple tumor antigens for treatment of patients with malignant melanoma: results from a phase I/II trial. Cytotherapy 12:721–734PubMedCrossRefGoogle Scholar
  32. 32.
    Lesterhuis WJ, Schreibelt G, Scharenborg NM et al (2011) Wild-type and modified gp100 peptide-pulsed dendritic cell vaccination of advanced melanoma patients can lead to long-term clinical responses independent of the peptide used. Cancer Immunol Immunother 60:249–260PubMedCrossRefGoogle Scholar
  33. 33.
    Nestle FO, Farkas A, Conrad C (2005) Dendritic-cell-based therapeutic vaccination against cancer. Curr Opin Immunol 17:163–169PubMedCrossRefGoogle Scholar
  34. 34.
    Radford KJ, Tullett KM, Lahoud MH (2014) Dendritic cells and cancer immunotherapy. Curr Opin Immunol 27:26–32PubMedCrossRefGoogle Scholar
  35. 35.
    Hodi FS, O’Day SJ, McDermott DF et al (2010) Improved survival with ipilimumab in patients with metastatic melanoma. N Engl J Med 363:711–723PubMedCentralPubMedCrossRefGoogle Scholar
  36. 36.
    Wolchok JD, Kluger H, Callahan MK et al (2013) Nivolumab plus Ipilimumab in Advanced Melanoma. N Engl J Med 369:122–133PubMedCrossRefGoogle Scholar
  37. 37.
    Maus MV, Thomas AK, Leonard DG, Allman D, Addya K, Schlienger K, Riley JL, June CH (2002) Ex vivo expansion of polyclonal and antigen-specific cytotoxic T lymphocytes by artificial APCs expressing ligands for the T-cell receptor, CD28 and 4-1BB. Nat Biotechnol 20:143–148PubMedCrossRefGoogle Scholar
  38. 38.
    Melero I, Bach N, Hellstrom KE, Aruffo A, Mittler RS, Chen L (1998) Amplification of tumor immunity by gene transfer of the co-stimulatory 4-1BB ligand: synergy with the CD28 co-stimulatory pathway. Eur J Immunol 28:1116–1121PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2015

Authors and Affiliations

  • Katie L. Powell
    • 1
    Email author
  • Alexandre S. Stephens
    • 1
  • Stephen J. Ralph
    • 1
  1. 1.School of Medical ScienceGriffith UniversityGold CoastAustralia

Personalised recommendations