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Replication-defective recombinant Semliki Forest virus encoding GM-CSF as a vector system for rapid and facile generation of autologous human tumor cell vaccines

Gene Therapy volume 8, pages 1515–1523 (2001)Cite this article

Abstract

This paper describes the production of recombinant Semliki Forest virus encoding murine or human granulocyte–macrophage colony-stimulating factor (GM-CSF) and the capacity of these vectors to transduce murine and human tumor cells ex vivo. High-titer stocks (up to 3 × 109 particles/ml) of conditionally infective, replication-defective, recombinant SFV particles were generated using the SFV Helper-2 system. It is shown that the recombinant SFV/GM-CSF virus, as well as recombinant SFV carrying the β-galactosidase reporter gene, efficiently transduce both murine tumor cell lines as well as primary human renal carcinoma cells. Using ELISA's specific for GM-CSF, levels of GM-CSF production by the cells were determined. Levels of murine GM-CSF (mGM-CSF) produced by SFV/mGM-CSF transduced renal cell cancer cultures were equal to or higher than corresponding levels reported in the literature after transduction of similar renal carcinoma cell cultures using a retroviral vector system. The biological activity of GM-CSF was demonstrated by using cells which are dependent on GM-CSF for growth and by using primary bone marrow cells. All the transduced cell cultures (including the human renal cell carcinoma samples) produced GM-CSF for up to at least 4 days after transduction. The results imply that the recombinant SFV system can be used for rapid and facile preparation of autologous cancer cell vaccines.

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Acknowledgements

The authors would like to thank P Liljeström and P Berglund for expert advice and valuable suggestions. In addition, we thank A Heikema, R Timmer, M Esselink and J Bijzet for their assistance with cloning, preparation of SFV batches and performing ELISAs. This work was funded by a grant from the Dutch Cancer Foundation (NKB/KWF).

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Author notes

  1. S Withoff and KL Glazenburg: SW and KLG contributed equally to this study

Authors and Affiliations

  1. Department of Medical Microbiology, Molecular Virology Section, University of Groningen and Academic Hospital, Groningen, The Netherlands

    S Withoff, KL Glazenburg, ML van Veen, MMJ Kraak, J Wilschut & T Daemen

  2. Department of Medical Oncology, University of Groningen and Academic Hospital, Groningen, The Netherlands

    GAP Hospers & EGE de Vries

  3. Institut für Pathologie, Universität Witten/Herdecke, Wuppertal, Germany

    S Störkel

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Withoff, S., Glazenburg, K., van Veen, M. et al. Replication-defective recombinant Semliki Forest virus encoding GM-CSF as a vector system for rapid and facile generation of autologous human tumor cell vaccines. Gene Ther 8, 1515–1523 (2001). https://doi.org/10.1038/sj.gt.3301556

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