Medical Oncology

, 31:191 | Cite as

Neoantigen in esophageal squamous cell carcinoma for dendritic cell-based cancer vaccine development

  • Mohammad Mahdi Forghanifard
  • Mehran Gholamin
  • Omeed Moaven
  • Moein Farshchian
  • Martha Ghahraman
  • Amir Aledavood
  • Mohammad Reza Abbaszadegan
Original Paper


Esophageal squamous cell carcinoma (ESCC) is a highly malignant tumor which usually is diagnosed in advanced stages due to its asymptomatic course of tumorigenesis. Current therapeutic modalities are not effective enough and the 5-year survival rate of the disease is still very low which prompts the urgent need for finding novel efficient therapeutic methods. In this study, we evaluated ex vivo immune response of ESCC patients against our newly designed chimeric construct consisting of highly immunogenic cancer-testis antigens. After confirming effective expression of the in vitro transcribed chimeric mRNA in ex vivo electroporated dendritic cells (DCs) of the ESCC patients, the patients’ CTLs were primed by DCs and cytotoxicity assay was performed to evaluate how the primed CTLs can recognize and target the chimeric mRNA-loaded cells. The chimeric protein was strongly expressed relative to the housekeeping gene expression in electroporated cells. The cytotoxicity of the CTLs was significantly higher in DCs loaded with chimeric mRNAs compared to mock DCs (p < 0.05) in all of the tested ESCC patients. We are introducing a novel construct that our functional study showed can stimulate and induce an effective immune response in ESCC patients. The designed chimeric mRNA-loaded DCs are capable of priming CTLs effectively and induce cytotoxicity against tumor. Therefore, loading DCs with chimeric epitopes of highly immunogenic antigens, such as cancer-testis antigens, are potentially interesting and effective therapeutic modalities for immunotherapy of ESCC.


Esophageal squamous cell carcinoma Immuno-gene therapy RNA vaccine CTAs Dendritic cell Chimeric construct Cytotoxicity 



The authors gratefully acknowledge the colleagues at the Departments of Surgery and Pathology in Omid, Qaem and Imam Reza hospitals of MUMS, Mashhad, for samples preparation. We are grateful to Dr. Mojtaba Sankian, Dr. Mohsen Tehrani and Ms. Maliheh Moghadam at Immunobiochemistry Lab, Immunology Research Center, Avicenna Research Institute, Mashhad, as well as our colleagues at Division of Human Genetics, for their kind help and support in technical aspects. This study was supported by the grant number 88098 from the Vice Chancellor for Research at MUMS.

Conflict of interest

The authors declare that they have no conflict of interest.


  1. 1.
    Jemal A, et al. Global patterns of cancer incidence and mortality rates and trends. Cancer Epidemiol Biomark Prev. 2010;19:1893–907.Google Scholar
  2. 2.
    The Japanese Society of Esophageal Diseases: Comprehensive registry of esophageal cancer in Japan (1998, 1999) and Long-term result of Esophagectomy in Japan (1988–1997), 2002.Google Scholar
  3. 3.
    Rice TW, et al. Worldwide esophageal cancer collaboration. Dis Esophagus. 2009;22:1–8.PubMedCrossRefGoogle Scholar
  4. 4.
    Milano F, Krishnadath KK. Novel therapeutic strategies for treating esophageal adenocarcinoma: the potential of dendritic cell immunotherapy and combinatorial regimens. Hum Immunol. 2008;69:614–24.PubMedCrossRefGoogle Scholar
  5. 5.
    Parish CR. Cancer immunotherapy: the past, the present and the future. Immunol Cell Biol. 2003;81:106–13.PubMedCrossRefGoogle Scholar
  6. 6.
    Smyth MJ, Godfrey DI, Trapani JA. A fresh look at tumor immunosurveillance and immunotherapy. Nat Immunol. 2001;2:293–9.PubMedCrossRefGoogle Scholar
  7. 7.
    Galluzzi L, et al. Trial watch: dendritic cell-based interventions for cancer therapy. Oncoimmunology 2012;1:1111–34.Google Scholar
  8. 8.
    Sahin U, Tureci et al. Expression of multiple cancer/testis (CT) antigens in breast cancer and melanoma: basis for polyvalent CT vaccine strategies. Int J Cancer. 1998;78:387–9.Google Scholar
  9. 9.
    Tajima K, et al. Expression of cancer/testis (CT) antigens in lung cancer. Lung Cancer. 2003;42:23–33.PubMedCrossRefGoogle Scholar
  10. 10.
    Forghanifard MM, et al. Cancer-testis gene expression profiling in esophageal squamous cell carcinoma: identification of specific tumor marker and potential targets for immunotherapy. Cancer Biol Ther. 2011;12:191–7.Google Scholar
  11. 11.
    Clark CE, Vonderheide RH. Cancer-testis antigens in tumor biology and immunotherapy. Cancer Biol Ther. 2006;5:1226–7.PubMedCrossRefGoogle Scholar
  12. 12.
    Forghanifard MM, et al. In silico analysis of chimeric polytope of cancer/testis antigens for dendritic cell-based immune-gene therapy applications. Gene Ther Mol Biol. 2012;14:87–96.Google Scholar
  13. 13.
    Gholamin M, et al. Induction of cytotoxic T lymphocytes primed with tumor RNA-loaded dendritic cells in esophageal squamous cell carcinoma: preliminary step for DC vaccine design. BMC Cancer. 2010;10:261.Google Scholar
  14. 14.
    Cheever MA, Higano CS. PROVENGE (Sipuleucel-T) in prostate cancer: the first FDA-approved therapeutic cancer vaccine. Clin Cancer Res. 2011;17:3520–6.Google Scholar
  15. 15.
    Palucka K, Banchereau J. Cancer immunotherapy via dendritic cells. Nat Rev Cancer. 2012;12:265–77.Google Scholar
  16. 16.
    Satoh Y, et al. Local administration of IL-12-transfected dendritic cells induces antitumor immune responses to colon adenocarcinoma in the liver in mice. J Exp Ther Oncol. 2002;2:337–49.PubMedCrossRefGoogle Scholar
  17. 17.
    Ueno H, et al. Harnessing human dendritic cell subsets for medicine. Immunol Rev. 2010;234:199–212.Google Scholar
  18. 18.
    Heiser A, et al. Autologous dendritic cells transfected with prostate-specific antigen RNA stimulate CTL responses against metastatic prostate tumors. J Clin Invest. 2002;109:409–17.PubMedCrossRefPubMedCentralGoogle Scholar
  19. 19.
    Koido S, et al. Induction of antitumor immunity by vaccination of dendritic cells transfected with MUC1 RNA. J Immunol. 2000;165:5713–9.PubMedCrossRefGoogle Scholar
  20. 20.
    Morse MA, et al. Immunotherapy with autologous, human dendritic cells transfected with carcinoembryonic antigen mRNA. Cancer Invest. 2003;21:341–9.PubMedCrossRefGoogle Scholar
  21. 21.
    Zeis M, et al. Generation of cytotoxic responses in mice and human individuals against hematological malignancies using survivin-RNA-transfected dendritic cells. J Immunol. 2003;170:5391–7.PubMedCrossRefGoogle Scholar
  22. 22.
    Tuting T, et al. Autologous human monocyte-derived dendritic cells genetically modified to express melanoma antigens elicit primary cytotoxic T cell responses in vitro: enhancement by cotransfection of genes encoding the Th1-biasing cytokines IL-12 and IFN-alpha. J Immunol. 1998;160:1139–47.PubMedGoogle Scholar
  23. 23.
    Tacken PJ, et al. Dendritic-cell immunotherapy: from ex vivo loading to in vivo targeting. Nat Rev Immunol. 2007;7:790–802.PubMedCrossRefGoogle Scholar
  24. 24.
    Van Brussel I, Berneman ZN, Cools N. Optimizing dendritic cell-based immunotherapy: tackling the complexity of different arms of the immune system. Mediators Inflamm. 2012. doi: 10.1155/2012/690643.
  25. 25.
    Lesterhuis WJ, Haanen JB, Punt CJ. Cancer immunotherapy–revisited. Nat Rev Drug Discov. 2011;10:591–600.Google Scholar
  26. 26.
    Rosenberg SA, Yang JC, Restifo NP. Cancer immunotherapy: moving beyond current vaccines. Nat Med. 2004;10:909–15.PubMedCrossRefPubMedCentralGoogle Scholar
  27. 27.
    Bujas T, et al. MAGE-A3/4 and NY-ESO-1 antigens expression in metastatic esophageal squamous cell carcinoma. Eur J Histochem. 2011;55:e7.Google Scholar
  28. 28.
    Quillien V, et al. Expression of MAGE genes in esophageal squamous-cell carcinoma. Anticancer Res. 1997;17:387–91.PubMedGoogle Scholar
  29. 29.
    Scanlan MJ, et al. Cancer/testis antigens: an expanding family of targets for cancer immunotherapy. Immunol Rev. 2002;188:22–32.PubMedCrossRefGoogle Scholar
  30. 30.
    Stevenson FK, Rice J, Zhu D. Tumor vaccines. Adv Immunol. 2004;82:49–103.PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  • Mohammad Mahdi Forghanifard
    • 1
  • Mehran Gholamin
    • 2
  • Omeed Moaven
    • 3
  • Moein Farshchian
    • 4
  • Martha Ghahraman
    • 5
  • Amir Aledavood
    • 6
  • Mohammad Reza Abbaszadegan
    • 2
    • 5
  1. 1.Department of Biology, Damghan BranchIslamic Azad UniversityDamghanIran
  2. 2.Division of Human Genetics, Immunology Research Center, Avicenna Research InstituteMashhad University of Medical SciencesMashhadIran
  3. 3.Department of Surgery, Massachusetts General HospitalHarvard Medical SchoolBostonUSA
  4. 4.Department of Biology, Faculty of ScienceFerdowsi University of MashhadMashhadIran
  5. 5.Medical Genetics Research Center, Medical SchoolMashhad University of Medical SciencesMashhadIran
  6. 6.Department of Radiotherapy and OncologyMashhad University of Medical ScienceMashhadIran

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