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Cancer Immunology, Immunotherapy

, Volume 63, Issue 3, pp 273–282 | Cite as

Enhanced therapeutic anti-tumor immunity induced by co-administration of 5-fluorouracil and adenovirus expressing CD40 ligand

  • Lina Liljenfeldt
  • Katerina Gkirtzimanaki
  • Dimitra Vyrla
  • Emma Svensson
  • Angelica SI Loskog
  • Aristides G. EliopoulosEmail author
Original Article

Abstract

Bystander immune activation by chemotherapy has recently gained extensive interest and provided support for the clinical use of chemotherapeutic agents in combination with immune enhancers. The CD40 ligand (CD40L; CD154) is a potent regulator of the anti-tumor immune response and recombinant adenovirus (RAd)-mediated CD40L gene therapy has been effective in various cancer models and in man. In this study we have assessed the combined effect of local RAd-CD40L and 5-fluorouracil (5-FU) administration on a syngeneic MB49 mouse bladder tumor model. Whereas MB49 cells implanted into immunocompetent mice responded poorly to RAd-CD40L or 5-FU alone, administration of both agents dramatically decreased tumor growth, increased survival of the mice and induced systemic MB49-specific immunity. This combination treatment was ineffective in athymic nude mice, highlighting an important role for T cell mediated anti-tumor immunity for full efficacy. 5-FU up-regulated the expression of Fas and immunogenic cell death markers in MB49 cells and cytotoxic T lymphocytes from mice receiving RAd-CD40L immunotherapy efficiently lysed 5-FU treated MB49 cells in a Fas ligand-dependent manner. Furthermore, local RAd-CD40L and 5-FU administration induced a shift of myeloid-derived suppressor cell phenotype into a less suppressive population. Collectively, these data suggest that RAd-CD40L gene therapy is a promising adjuvant treatment to 5-FU for the management of bladder cancer.

Keywords

CD40L 5-Fluorouracil Immunotherapy Chemotherapy Urinary bladder cancer 

Notes

Acknowledgments

The authors thank Berith Nilsson at Uppsala University for technical assistance with viral vector production and Marina Ioannou at IMBB-FORTH for cell culture assistance. This work was supported by the European Commission FP6 program Apotherapy (EC contract number 037344) to Aristides Eliopoulos and Angelica Loskog, the EC FP7 programmes INFLA-CARE (EC contract number 223151) and ‘Translational Potential’ (TransPOT; EC contract number 285948) to Aristides Eliopoulos, and by a Swedish Research Council grant to Angelica Loskog. Aristides Eliopoulos also acknowledges co-funding of this research by the General Secretariat of Research and Technology of Greece through the Operational Program Competitiveness and Entrepreneurship (OPC II), NSRF 2007-2013, action “SYNERGASIA 2011”, Project THERA-CAN (contract number 11ΣΥΝ_1_485).

Conflict of interest

The authors have no conflicts of interest to declare except from Angelica Loskog who is the CEO of Lokon Pharma AB, a scientific advisor at NEXTTOBE AB and has a royalty agreement with Alligator Bioscience AB.

Supplementary material

262_2013_1507_MOESM1_ESM.pdf (89 kb)
Supplementary material 1 (PDF 89 kb)

References

  1. 1.
    van Kooten C, Banchereau J (2000) CD40-CD40 ligand. J Leukoc Biol 67:2–17PubMedGoogle Scholar
  2. 2.
    Loskog AS, Eliopoulos AG (2009) The Janus faces of CD40 in cancer. Semin Immunol 21:301–307PubMedCrossRefGoogle Scholar
  3. 3.
    Callard RE, Armitage RJ, Fanslow WC, Spriggs MK (1993) CD40 ligand and its role in X-linked hyper-IgM syndrome. Immunol Today 14:559–564PubMedCrossRefGoogle Scholar
  4. 4.
    Hayward AR, Levy J, Facchetti F, Notarangelo L, Ochs HD, Etzioni A, Bonnefoy JY, Cosyns M, Weinberg A (1997) Cholangiopathy and tumors of the pancreas, liver, and biliary tree in boys with X-linked immunodeficiency with hyper-IgM. J Immunol 158:977–983PubMedGoogle Scholar
  5. 5.
    Mackey MF, Gunn JR, Maliszewsky C, Kikutani H, Noelle RJ, Barth RJ Jr (1998) Dendritic cells require maturation via CD40 to generate protective antitumor immunity. J Immunol 161:2094–2098PubMedGoogle Scholar
  6. 6.
    French RR, Chan HT, Tutt AL, Glennie MJ (1999) CD40 antibody evokes a cytotoxic T-cell response that eradicates lymphoma and bypasses T-cell help. Nat Med 5:548–553PubMedCrossRefGoogle Scholar
  7. 7.
    Kikuchi T, Crystal RG (1999) Anti-tumor immunity induced by in vivo adenovirus vector-mediated expression of CD40 ligand in tumor cells. Hum Gene Ther 10:1375–1387PubMedCrossRefGoogle Scholar
  8. 8.
    Loskog A, Bjorkland A, Brown MP, Korsgren O, Malmstrom PU, Totterman TH (2001) Potent antitumor effects of CD154 transduced tumor cells in experimental bladder cancer. J Urol 166:1093–1097PubMedCrossRefGoogle Scholar
  9. 9.
    Noguchi M, Imaizumi K, Kawabe T et al (2001) Induction of antitumor immunity by transduction of CD40 ligand gene and interferon-gamma gene into lung cancer. Cancer Gene Ther 8:421–429PubMedCrossRefGoogle Scholar
  10. 10.
    Todryk SM, Tutt AL, Green MH, Smallwood JA, Halanek N, Dalgleish AG, Glennie MJ (2001) CD40 ligation for immunotherapy of solid tumours. J Immunol Methods 248:139–147PubMedCrossRefGoogle Scholar
  11. 11.
    Dzojic H, Loskog A, Totterman TH, Essand M (2006) Adenovirus-mediated CD40 ligand therapy induces tumor cell apoptosis and systemic immunity in the TRAMP-C2 mouse prostate cancer model. Prostate 66:831–838PubMedCrossRefGoogle Scholar
  12. 12.
    Sun Y, Peng D, Lecanda J, Schmitz V, Barajas M, Qian C, Prieto J (2000) In vivo gene transfer of CD40 ligand into colon cancer cells induces local production of cytokines and chemokines, tumor eradication and protective antitumor immunity. Gene Ther 7:1467–1476PubMedCrossRefGoogle Scholar
  13. 13.
    Buhtoiarov IN, Lum H, Berke G, Paulnock DM, Sondel PM, Rakhmilevich AL (2005) CD40 ligation activates murine macrophages via an IFN-gamma-dependent mechanism resulting in tumor cell destruction in vitro. J Immunol 174:6013–6022PubMedGoogle Scholar
  14. 14.
    Sabel MS, Yamada M, Kawaguchi Y, Chen FA, Takita H, Bankert RB (2000) CD40 expression on human lung cancer correlates with metastatic spread. Cancer Immunol Immunother 49:101–108PubMedCrossRefGoogle Scholar
  15. 15.
    Hirano A, Longo DL, Taub DD et al (1999) Inhibition of human breast carcinoma growth by a soluble recombinant human CD40 ligand. Blood 93:2999–3007PubMedGoogle Scholar
  16. 16.
    Tong AW, Papayoti MH, Netto G, Armstrong DT, Ordonez G, Lawson JM, Stone MJ (2001) Growth-inhibitory effects of CD40 ligand (CD154) and its endogenous expression in human breast cancer. Clin Cancer Res 7:691–703PubMedGoogle Scholar
  17. 17.
    Eliopoulos AG, Dawson CW, Mosialos G et al (1996) CD40-induced growth inhibition in epithelial cells is mimicked by Epstein-Barr Virus-encoded LMP1: involvement of TRAF3 as a common mediator. Oncogene 13:2243–2254PubMedGoogle Scholar
  18. 18.
    Knox PG, Davies CC, Ioannou M, Eliopoulos AG (2011) The death domain kinase RIP1 links the immunoregulatory CD40 receptor to apoptotic signaling in carcinomas. J Cell Biol 192:391–399PubMedCrossRefGoogle Scholar
  19. 19.
    Afford SC, Randhawa S, Eliopoulos AG, Hubscher SG, Young LS, Adams DH (1999) CD40 activation induces apoptosis in cultured human hepatocytes via induction of cell surface fas ligand expression and amplifies fas-mediated hepatocyte death during allograft rejection. J Exp Med 189:441–446PubMedCentralPubMedCrossRefGoogle Scholar
  20. 20.
    Ghamande S, Hylander BL, Oflazoglu E, Lele S, Fanslow W, Repasky EA (2001) Recombinant CD40 ligand therapy has significant antitumor effects on CD40-positive ovarian tumor xenografts grown in SCID mice and demonstrates an augmented effect with cisplatin. Cancer Res 61:7556–7562PubMedGoogle Scholar
  21. 21.
    Moschonas A, Kouraki M, Knox PG, Thymiakou E, Kardassis D, Eliopoulos AG (2008) CD40 induces antigen transporter and immunoproteasome gene expression in carcinomas via the coordinated action of NF-kappaB and of NF-kappaB-mediated de novo synthesis of IRF-1. Mol Cell Biol 28:6208–6222PubMedCentralPubMedCrossRefGoogle Scholar
  22. 22.
    Gomes EM, Rodrigues MS, Phadke AP et al (2009) Antitumor activity of an oncolytic adenoviral-CD40 ligand (CD154) transgene construct in human breast cancer cells. Clin Cancer Res 15:1317–1325PubMedCrossRefGoogle Scholar
  23. 23.
    Diaconu I, Cerullo V, Hirvinen ML et al (2012) Immune response is an important aspect of the antitumor effect produced by a CD40L-encoding oncolytic adenovirus. Cancer Res 72:2327–2338PubMedCrossRefGoogle Scholar
  24. 24.
    Vonderheide RH, Butler MO, Liu JF, Battle TE, Hirano N, Gribben JG, Frank DA, Schultze JL, Nadler LM (2001) CD40 activation of carcinoma cells increases expression of adhesion and major histocompatibility molecules but fails to induce either CD80/CD86 expression or T cell alloreactivity. Int J Oncol 19:791–798PubMedGoogle Scholar
  25. 25.
    Malmstrom PU, Loskog AS, Lindqvist CA, Mangsbo SM, Fransson M, Wanders A, Gardmark T, Totterman TH (2010) AdCD40L immunogene therapy for bladder carcinoma: the first phase I/IIa trial. Clin Cancer Res 16:3279–3287PubMedCrossRefGoogle Scholar
  26. 26.
    Beatty GL, Chiorean EG, Fishman MP et al (2011) CD40 agonists alter tumor stroma and show efficacy against pancreatic carcinoma in mice and humans. Science 331:1612–1616PubMedCentralPubMedCrossRefGoogle Scholar
  27. 27.
    Pesonen S, Diaconu I, Kangasniemi L et al (2012) Oncolytic immunotherapy of advanced solid tumors with a CD40L-expressing replicating adenovirus: assessment of safety and immunologic responses in patients. Cancer Res 72:1621–1631PubMedCrossRefGoogle Scholar
  28. 28.
    Folkman J, Hahnfeldt P, Hlatky L (2000) Cancer: looking outside the genome. Nat Rev Mol Cell Biol 1:76–79PubMedCrossRefGoogle Scholar
  29. 29.
    Loskog A, Dzojic H, Vikman S, Ninalga C, Essand M, Korsgren O, Totterman TH (2004) Adenovirus CD40 ligand gene therapy counteracts immune escape mechanisms in the tumor microenvironment. J Immunol 172:7200–7205PubMedGoogle Scholar
  30. 30.
    Jackaman C, Nelson DJ (2012) Intratumoral interleukin-2/agonist CD40 antibody drives CD4+ -independent resolution of treated-tumors and CD4+ -dependent systemic and memory responses. Cancer Immunol Immunother 61:549–560PubMedCrossRefGoogle Scholar
  31. 31.
    Loskog AS, Fransson ME, Totterman TT (2005) AdCD40L gene therapy counteracts T regulatory cells and cures aggressive tumors in an orthotopic bladder cancer model. Clin Cancer Res 11:8816–8821PubMedCrossRefGoogle Scholar
  32. 32.
    Eliopoulos AG, Wang CC, Dumitru CD, Tsichlis PN (2003) Tpl2 transduces CD40 and TNF signals that activate ERK and regulates IgE induction by CD40. EMBO J 22:3855–3864PubMedCrossRefGoogle Scholar
  33. 33.
    Hannani D, Sistigu A, Kepp O, Galluzzi L, Kroemer G, Zitvogel L (2011) Prerequisites for the antitumor vaccine-like effect of chemotherapy and radiotherapy. Cancer J 17:351–358PubMedCrossRefGoogle Scholar
  34. 34.
    Gabrilovich DI, Nagaraj S (2009) Myeloid-derived suppressor cells as regulators of the immune system. Nat Rev Immunol 9:162–174PubMedCentralPubMedCrossRefGoogle Scholar
  35. 35.
    Dolcetti L, Peranzoni E, Ugel S et al (2010) Hierarchy of immunosuppressive strength among myeloid-derived suppressor cell subsets is determined by GM-CSF. Eur J Immunol 40:22–35PubMedCrossRefGoogle Scholar
  36. 36.
    Apetoh L, Ghiringhelli F, Tesniere A et al (2007) Toll-like receptor 4-dependent contribution of the immune system to anticancer chemotherapy and radiotherapy. Nat Med 13:1050–1059PubMedCrossRefGoogle Scholar
  37. 37.
    Tesniere A, Schlemmer F, Boige V et al (2010) Immunogenic death of colon cancer cells treated with oxaliplatin. Oncogene 29:482–491PubMedCrossRefGoogle Scholar
  38. 38.
    Ramakrishnan R, Gabrilovich DI (2013) Novel mechanism of synergistic effects of conventional chemotherapy and immune therapy of cancer. Cancer Immunol Immunother 62:405–410PubMedCrossRefGoogle Scholar
  39. 39.
    Zitvogel L, Kepp O, Kroemer G (2011) Immune parameters affecting the efficacy of chemotherapeutic regimens. Nat Rev Clin Oncol 8:151–160PubMedCrossRefGoogle Scholar
  40. 40.
    Nowak AK, Robinson BW, Lake RA (2003) Synergy between chemotherapy and immunotherapy in the treatment of established murine solid tumors. Cancer Res 63:4490–4496PubMedGoogle Scholar
  41. 41.
    Vardouli L, Lindqvist C, Vlahou K, Loskog AS, Eliopoulos AG (2009) Adenovirus delivery of human CD40 ligand gene confers direct therapeutic effects on carcinomas. Cancer Gene Ther 16:848–860PubMedCrossRefGoogle Scholar
  42. 42.
    Vincent J, Mignot G, Chalmin F et al (2010) 5-Fluorouracil selectively kills tumor-associated myeloid-derived suppressor cells resulting in enhanced T cell-dependent antitumor immunity. Cancer Res 70:3052–3061PubMedCrossRefGoogle Scholar
  43. 43.
    Stewart TJ, Abrams SI (2008) How tumours escape mass destruction. Oncogene 27:5894–5903PubMedCrossRefGoogle Scholar
  44. 44.
    Yeh KY, Pulaski BA, Woods ML, McAdam AJ, Gaspari AA, Frelinger JG, Lord EM (1995) B7-1 enhances natural killer cell-mediated cytotoxicity and inhibits tumor growth of a poorly immunogenic murine carcinoma. Cell Immunol 165:217–224PubMedCrossRefGoogle Scholar
  45. 45.
    Cooke PW, James ND, Ganesan R, Wallace M, Burton A, Young LS (1999) CD40 expression in bladder cancer. J Pathol 188:38–43PubMedCrossRefGoogle Scholar
  46. 46.
    Bugajska U, Georgopoulos NT, Southgate J, Johnson PW, Graber P, Gordon J, Selby PJ, Trejdosiewicz LK (2002) The effects of malignant transformation on susceptibility of human urothelial cells to CD40-mediated apoptosis. J Natl Cancer Inst 94:1381–1395PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2013

Authors and Affiliations

  • Lina Liljenfeldt
    • 1
  • Katerina Gkirtzimanaki
    • 2
    • 3
  • Dimitra Vyrla
    • 2
    • 3
  • Emma Svensson
    • 1
  • Angelica SI Loskog
    • 1
  • Aristides G. Eliopoulos
    • 2
    • 3
    • 4
    Email author
  1. 1.Science for Life Laboratory, Department of Immunology, Genetics and PathologyUppsala UniversityUppsalaSweden
  2. 2.Molecular and Cellular Biology Laboratory, Division of Basic SciencesUniversity of Crete Medical SchoolHeraklion, CreteGreece
  3. 3.Laboratory of Cancer Biology, Institute of Molecular Biology and BiotechnologyFORTHHeraklion, CreteGreece
  4. 4.Laboratory of Translational Medicine and Experimental TherapeuticsUniversity of Crete Medical SchoolHeraklionGreece

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