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Transfer of mRNA encoding recombinant immunoreceptors reprograms CD4+ and CD8+ T cells for use in the adoptive immunotherapy of cancer

Gene Therapy volume 16, pages 596–604 (2009)Cite this article

Abstract

Human T lymphocytes can be redirected with a new defined specificity by expression of a chimeric T-cell receptor (immunoreceptor) for the use in adoptive immunotherapy of cancer. Whereas standard procedures use retroviral gene transduction to constitutively express immunoreceptors in T cells, we here explored for the first time mRNA electroporation to achieve transient immunoreceptor expression, and thereby minimizing the risk of persistence of potential autoaggression. CD4+ and CD8+ T cells were efficiently transfected with immunoreceptors specific for ErbB2 and CEA. The immunoreceptor expression was transient with half-maximal expression at day 2 and no detectable immunoreceptor expression at day 9 after electroporation. Immunoreceptor-transfected T cells were specifically activated upon coincubation with ErbB2+ and CEA+ tumor cells, respectively, resulting in secretion of interferon-γ (IFNγ), interleukin-2 (IL-2), and tumor necrosis factor-α (TNFα). Furthermore, immunoreceptor-transfected CD8+ T cells specifically lysed ErbB2+ and CEA+ tumor cells, respectively. The RNA-transfected T cells retained their cytotoxic function after 2 days of activation and exhibited cytolytic activities like retrovirally transduced T cells. RNA electroporation of T cells thereby provides a versatile tool for transient immunoreceptor expression, which may be of advantage in avoiding the persistence of unintended autoaggression.

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Acknowledgements

We thank Tanja de Gruijl and Saskia Santegoets (VU University Medical Center, Amsterdam) for providing us with the KATO III cell line, Kris Thielemans (Medical School of the Vrije Universiteit Brussel, Brussels) for providing the pGEM4Z-5′UTR-sig-MAGE-A3-DC.LAMP-3′UTR RNA production vector. We thank Tanja Schunder, Verena Wellner, and Stefanie Baumann for excellent technical assistance. We thank Stefanie Hoyer and Christian Hofmann for fruitful discussions. Work in the Erlangen laboratory was financially supported by the ELAN-Funds of the Friedrich-Alexander-University Erlangen-Nuremberg (DE-07.09.12.1) and the DFG-German Research Foundation (SCHA 1247/1-1). Work in the Cologne laboratory was supported by the European Community through ATTACK and by the Fortune program of the Medical Faculty.

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Author notes

  1. N Schaft and J Dörrie: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Dermatology, University Hospital Erlangen, Erlangen, Germany

    K Birkholz, C Krug, S Reuter, E Kämpgen, G Schuler, N Schaft & J Dörrie

  2. Center for Molecular Medicine Cologne (CMMC), University of Cologne, Cologne, Germany

    A Hombach & H Abken

  3. Cancer Immunology Section, Peter MacCallum Cancer Centre, East Melbourne, Victoria, Australia

    M Kershaw

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  1. K Birkholz
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Correspondence to N Schaft.

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Birkholz, K., Hombach, A., Krug, C. et al. Transfer of mRNA encoding recombinant immunoreceptors reprograms CD4+ and CD8+ T cells for use in the adoptive immunotherapy of cancer. Gene Ther 16, 596–604 (2009). https://doi.org/10.1038/gt.2008.189

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