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Novel adenovirus-based helper system to support production of recombinant parvovirus

Cancer Gene Therapy volume 18, pages 240–249 (2011)Cite this article

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Abstract

Preclinical studies using various cell culture and animal systems highlight the potential of recombinant rodent parvoviruses (recPVs) for cancer therapy. Production of these viruses is, however, not efficient and this hampers the clinical applications of these agents. In this study, we show that the adenovirus genes E2a, E4(orf6) and VA RNA increase the production of recPVs by more than 10-fold and reduce the time of production from 3 to 2 days in HEK293T cells. The helper effects of these genes can be observed with different recPVs, regardless of the nature and size of the inserted transgene. Furthermore, we generated a recombinant Adenovirus 5 carrying the parvovirus VP transcription unit. This helper, named Ad-VP, allows recPVs to be efficiently produced through a protocol based only on cell infection, making possible to use cell lines, such as NB324K, which are good producers of parvoviruses but are hardly transfectable. Hence, we could further improve viral titers and reduce time and costs of production. This Ad-VP helper-based protocol could be scaled up to a bioreactor format for the generation of the large amounts of recPVs needed for future clinical applications.

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Acknowledgements

We thank the team of the DKFZ virus production and development unit, in particular Markus Müller, Silvia Münstermann and Barbara Liebertrau for their valuable help with virus production and characterization. We are also indebted to Melanie Krämer for technical assistance and Nathalie Salomé and Michele Vogel for the anti-NS1 antibody. NEA was supported by long-term fellowships from Alexander von Humboldt-Foundation, (Bonn, Germany) and the European Molecular Biology Organization (Heidelberg, Germany). This work was supported by a grant from the Helmholtz-Gemeinschaft in the frame of the Deutsches Krebsforschungszentrum/Cancéropôle du Grand-Est joint Programme in Applied Tumor Virology.

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Authors and Affiliations

  1. Infection and Cancer Program, German Cancer Research Center (DKFZ), Heidelberg, Germany

    N El-Andaloussi, M Endele, B Leuchs, S Bonifati, J Kleinschmidt, J Rommelaere & A Marchini

  2. Inserm U701, Heidelberg, Germany

    N El-Andaloussi, J Rommelaere & A Marchini

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  1. N El-Andaloussi
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  2. M Endele
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  3. B Leuchs
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Correspondence to A Marchini.

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A patent application, based on the results described in this article, has been submitted by the authors to the European Patent Office.

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El-Andaloussi, N., Endele, M., Leuchs, B. et al. Novel adenovirus-based helper system to support production of recombinant parvovirus. Cancer Gene Ther 18, 240–249 (2011). https://doi.org/10.1038/cgt.2010.73

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