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Induction of antigen-specific CD8+ cytotoxic T cells by dendritic cells co-electroporated with a dsRNA analogue and tumor antigen mRNA

Gene Therapy volume 13, pages 1027–1036 (2006)Cite this article

Abstract

The maturation state of dendritic cells (DCs) is an important determinant for the initiation and regulation of adaptive immune responses. In this study, we wanted to assess whether functional activation of human monocyte-derived DCs can be achieved by electroporation of an activation signal in the form of double-stranded (ds) RNA and whether simultaneous electroporation of the dsRNA with tumor antigen encoding mRNA can lead to the induction of a cytotoxic T-lymphocyte (CTL) response. Electroporation of immature DCs with poly(I:C12U), a dsRNA analogue, resulted in phenotypic as well as functional changes, indicative of DC maturation. Co-electroporation of DCs with both poly(I:C12U) and Melan-A/MART-1 encoding mRNA induced strong anti-Melan-A/MART-1 CD8+ T-cell responses in vitro. Higher numbers of Melan-A/MART-1-specific CTLs were consistently obtained with poly(I:C12U)-activated DCs compared to DCs matured in the presence of an inflammatory cytokine cocktail. These results indicate that DC co-electroporation with both dsRNA and tumor antigen encoding mRNA induces fully activated and antigen-loaded DCs that promote antigen-specific CTL responses and may provide the basis for future immunotherapeutic strategies.

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Acknowledgements

We thank Erna Borms, Christine Huysmans and Jan Volkaert for help with DC cultures, Elsy Vaeremans and Peggy Verbuyst for mRNA preparations, Roger Andries and Jos Theunissen for their helpful technical suggestions and Daniel Powell and Gregory Lizée for critical reading of the manuscript. This work was supported by grants to Kris Thielemans from the Fund for Scientific Research-Flanders (FWO-Vlaanderen), the Ministry of Science (IUAP/PAI IV), the Fortis Bank, the Belgische Federatie voor Kankerbestrijding and BruCells. JL Aerts was supported by a return grant from the Belgian Science Policy (BELSPO).

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  1. Department of Physiology and Immunology, Laboratory of Molecular and Cellular Therapy, Medical School of the Vrije Universiteit Brussel (VUB), Brussels, Belgium

    A Michiels, K Breckpot, J Corthals, S Tuyaerts, A Bonehill, C Heirman, K Thielemans & J L Aerts

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Correspondence to J L Aerts.

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Michiels, A., Breckpot, K., Corthals, J. et al. Induction of antigen-specific CD8+ cytotoxic T cells by dendritic cells co-electroporated with a dsRNA analogue and tumor antigen mRNA. Gene Ther 13, 1027–1036 (2006). https://doi.org/10.1038/sj.gt.3302750

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