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Induction of Influenza Matrix Protein 1 and MelanA-specific T lymphocytes in vitro using mRNA-electroporated dendritic cells

Cancer Gene Therapy volume 10, pages 696–706 (2003)Cite this article

Abstract

Genetically modified dendritic cells (DC) constitute a promising approach in cancer immunotherapy. Viral gene delivery systems have been shown to be very efficient strategies, but safety concerns for their clinical use in immunotherapy remain an important issue. Recently, the technique of mRNA electroporation was described as a very efficient tool for the genetic modification of human monocyte-derived DC. Here, we show that transgene expression can be modulated by varying the amount of mRNA used for electroporation. We document that CD40 ligation leads to a significant production of IL-12 by the electroporated DC, although the level of IL-12 production is somewhat lower than for non- or mock-electroporated DC. Furthermore, we show that the electroporated DC can be frozen and thawed without loss of viability or function and that Influenza virus Matrix Protein 1 mRNA electroporated DC are capable of inducing a memory cytotoxic T lymphocyte response and are more potent in doing so than mRNA-pulsed DC. Similar results were obtained with MelanA/MART-1 mRNA electroporated DC. These results clearly indicate that mRNA-electroporated DC represent powerful candidates for use as tumor vaccines and could constitute an improvement compared with vaccines using peptide-pulsed DC.

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Acknowledgements

We thank Christine Huysmans, Danny Carels and Katja Michielsen for the help with DC cultures, Elsy Vaeremans and Peggy Verbuyst for the mRNA preparation, Karolien Claes for performing IFNγ-ELISPOT assays and Jos Theunissen for useful discussions.

This work was supported by grants to K T from the Fund for Scientific Research-Flanders (FWO-Vlaanderen), the Institute for Science & Technology (IWT), the Ministry of Science (IUAP/PAI IV), the FORTIS Bank and De Belgische Federatie voor Kankerbestrijding.

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  1. Department of Physiology–Immunology, Laboratory of Molecular and Cellular Therapy, Medical School of the Vrije Universiteit Brussel (VUB), Laarbeeklaan 103/E, Brussels, 1090, Belgium

    Sandra Tuyaerts, Annelies Michiels, Jurgen Corthals, Aude Bonehill, Carlo Heirman, Catherine De Greef, Sofie M Noppe & Kris Thielemans

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Correspondence to Kris Thielemans.

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Tuyaerts, S., Michiels, A., Corthals, J. et al. Induction of Influenza Matrix Protein 1 and MelanA-specific T lymphocytes in vitro using mRNA-electroporated dendritic cells. Cancer Gene Ther 10, 696–706 (2003). https://doi.org/10.1038/sj.cgt.7700622

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