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


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.


CD40L 5-Fluorouracil Immunotherapy Chemotherapy Urinary bladder cancer 



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)


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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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