Dendritic Cell Vaccines for Cancer Stem Cells

  • Serena Pellegatta
  • Gaetano Finocchiaro
Part of the Methods in Molecular Biology book series (MIMB, volume 568)


Accumulating evidence suggests that only a fraction of neoplastic cells, defined as cancer stem cells (CSC), are responsible for tumor perpetuation. Recent data suggest that neurospheres (NS) from glioblastoma multiforme (GBM) are enriched in CSC. The characterization of this subpopulation of brain tumor cells with a potent tumorigenic activity supports the cancer stem cell hypothesis in solid tumors and may imply that cancer cells are differentially targeted by treatments, including dendritic cell (DC) immunotherapy. To test therapeutic strategies, a good model mimicking the characteristics of GBM-NS and GBM-AC (Adherent Cells) was necessary. One of the most frequently used murine brain tumor models is the GL261 glioma cell line. To see whether GL261 cells could mimic the growth of human GBM-CSC we let them grow in EGF/bFGF without serum. After 5 days neurospheres were visible in the culture medium and were proliferating continuously. The characterization in vivo and in vitro demonstrates that GL261-NS satisfy criteria used to identify CSC and are more immunogenic than AC. DC loaded with GL261-NS lysates protect mice against tumors from both GL261-NS and GL261-AC. Our results suggest that only DC vaccination against neurospheres can restrain the growth of a highly infiltrating and aggressive model of glioma and may have implications for the design of novel, more effective immunotherapy trials for malignant glioma and possibly other malignancies.

Key words

Dendritic cells Cancer stem cells GL261 glioma cells Neurospheres Glioblastoma multiforme 



We acknowledge the support of AIRC, ARIN-Fondo di Gio, and Italian Minister of Health.


  1. 1.
    Holland, E. C. (2000) Glioblastoma multiforme: the terminator. Proc Natl Acad Sci U S A. 97, 6242–6244.CrossRefGoogle Scholar
  2. 2.
    Castro, M. G., Cowen, R., Williamson, I. K., David, A., Jimenez-Dalmaroni, M. J., Yuan, X., Bigliari, A., Williams, J. C., Hu, J., and Lowenstein, P. R. (2003) Current and future strategies for the treatment of malignant brain tumors. Pharmacol Ther. 98, 71–108.PubMedCrossRefGoogle Scholar
  3. 3.
    Stupp, R., Mason, W. P., Van Den Bent, M. J., Weller, M., Fisher, B., Taphoorn, M., Brandes, A. A., Cairncross, G., Lacombe, D., and Mirimanoff, R. O. Concomitant and adjuvant temozolomide (TMZ) and radiotherapy(RT) for newly diagnosed glioblastoma multiforme (GBM) (2004). Conclusive results of a randomized phase III trial by the EORTC Brain & RT Groups and NCIC Clinical Trials Group. J Clin Oncol (Meeting Abstracts). 22, 2.Google Scholar
  4. 4.
    Ehtesham, M., Black, K. L., and Yu, J. S. (2004) Recent progress in immunotherapy for malignant glioma: treatment strategies and results from clinical trials. Cancer Control 11, 192–207.PubMedGoogle Scholar
  5. 5.
    Yu, J. S., Wheeler, C. J., Zeltzer, P. M., Ying, H., Finger, D. N., Lee, P. K., Yong, W. H., Incardona, F., Thompson, R. C., Riedinger, M. S., Zhang, W., Prins, R. M., and Black, K. L. (2001) Vaccination of malignant glioma patients with peptide-pulsed dendritic cells elicits systemic cytotoxicity and intracranial T-cell infiltration. Cancer Res. 61, 842–847.PubMedGoogle Scholar
  6. 6.
    Yamanaka, R., Abe, T., Yajima, N., Tsuchiya, N., Homma, J., Kobayashi, T., Narita, M., Takahashi, M., and Tanaka, R. (2003) Vaccination of recurrent glioma patients with tumour lysate-pulsed dendritic cells elicits immune responses: results of a clinical phase I/II trial. Br J Cancer. 89, 1172–1179.PubMedCrossRefGoogle Scholar
  7. 7.
    Rutkowski, S., De Vleeschouwer, S., Kaempgen, E., Wolff, J. E., Kuhl, J., Demaerel, P., Warmuth-Metz, M., Flamen, P., Van Calenbergh, F., Plets, C., Sorensen, N., Opitz, A., and Van Gool, S. W. (2004) Surgery and adjuvant dendritic cell-based tumour vaccination for patients with relapsed malignant glioma, a feasibility study. Br J Cancer. 91, 1656–1662.PubMedGoogle Scholar
  8. 8.
    Yu, J. S., Liu, G., Ying, H., Yong, W. H., Black, K. L., and Wheeler, C. J. (2004) Vaccination with tumor lysate-pulsed dendritic cells elicits antigen-specific, cytotoxic T-cells in patients with malignant glioma. Cancer Res. 64, 4973–4979.PubMedCrossRefGoogle Scholar
  9. 9.
    Yamanaka, R., Homma, J., Yajima, N., Tsuchiya, N., Sano, M., Kobayashi, T., Yoshida, S., Abe, T., Narita, M., Takahashi, M., et al. (2005) Clinical evaluation of dendritic cell vaccination for patients with recurrent glioma: results of a clinical phase I/II trial. Clin Cancer Res. 11, 4160–4167.PubMedCrossRefGoogle Scholar
  10. 10.
    Clarke, M. F. (2004) Neurobiology: at the root of brain cancer. Nature. 432, 281–282.PubMedCrossRefGoogle Scholar
  11. 11.
    Al-Hajj, M., Becker, M. W., Wicha, M., Weissman, I., and Clarke, M. F. (2004) Therapeutic implications of cancer stem cells. Curr Opin Genet Dev. 14, 43–47.PubMedCrossRefGoogle Scholar
  12. 12.
    Pellegatta, S., Poliani, P.L., Corno, D., Menghi, F., Ghielmetti, F., Suarez-Merino, B., Caldera, V., Nava, S., Ravanini, M., Facchetti, F., Bruzzone, MG., Finocchiaroan G. (2006) Neurospheres enriched in cancer stem-like cells are highly effective in eliciting a dendritic cell-mediated immune response against malignant gliomas. Cancer Res. 66, 10247–10252.PubMedCrossRefGoogle Scholar
  13. 13.
    Ausman, J.I., Shapiro, W.R., & Rall, D.P. (1970). Studies on the chemotherapy of experimental brain tumors: development of an experimental model. Cancer Res 30, 2394–2400.PubMedGoogle Scholar
  14. 14.
    Benedetti, S., Pirola, B., Pollo, B., Magrassi, L., Bruzzone, M. G., Rigamonti, D., Galli, R., Selleri, S., Di Meco, F., De Fraja, C., Vescovi, A., Cattaneo, E., and Finocchiaro, G. (2000) Gene therapy of experimental brain tumors using neural progenitor cells. Nat Med 6, 447–450.PubMedCrossRefGoogle Scholar
  15. 15.
    Plautz, G.E., Touhalisky, J.E., & Shu, S. (1997). Treatment of murine gliomas by adoptive transfer of ex vivo activated tumor-draining lymph node cells. Cell Immunol. 178,101–107.PubMedCrossRefGoogle Scholar
  16. 16.
    Vescovi, A.L., Galli, R., Reynolds, B.A. (2006) Brain tumor stem cells. Nat.Rev. Canc. 6, 425–436.CrossRefGoogle Scholar

Copyright information

© Humana Press, a part of Springer Science+Business Media, LLC 2009

Authors and Affiliations

  • Serena Pellegatta
    • 1
  • Gaetano Finocchiaro
    • 1
  1. 1.Unit of Experimental Neuro-OncologyFondazione IRCCS Istituto Neurologico BestaMilanoItaly

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