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Cancer Vaccines pp 3–15Cite as

Single-Step Antigen Loading and Maturation of Dendritic Cells Through mRNA Electroporation of a Tumor-Associated Antigen and a TriMix of Costimulatory Molecules

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1139))

Abstract

Dendritic cells (DC) are key players in several types of cancer vaccines. Large numbers of DC can easily be generated in closed systems from the monocyte fraction of the peripheral blood. They are the professional antigen-presenting cells, and electroporation of mRNA-encoding tumor antigens is a very efficient and a relatively simple way to load the DC with antigen. The co-electroporation of a tumor antigen of choice and the combination of 3 costimulatory molecules, including CD70, caTLR4, and CD40L (TriMix-DC), leads to fully potent antigen-presenting DC able to generate a broad immune response.

Here we describe the in vitro transcription of the mRNA and the subsequent generation and electroporation of autologous DC used for the treatment of melanoma patients.

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Acknowledgements

TriMix-DC are the topic of a current patent application (WO2009/034172). A. B. and K. T. are mentioned as inventors of this application. None of the authors involved in this study receives any form of support or remuneration related to this platform.

This work was supported by grants from the Interuniversity Attraction Poles Program–Belgian State–Belgian Science Policy, the National Cancer Plan of the Federal Ministry of Health, the Stichting tegen Kanker, the Vlaamse Liga tegen Kanker, an Integrated Project and a Network of Excellence sponsored by the EU FP-6, an IWT-TBM program, the Fonds voor Wetenschappelijk Onderzoek Vlaanderen (FWO-Vlaanderen), and the Willy Gepts Wetenschappelijk Fonds of the UZ Brussel. S. W. is a Ph.D. fellow and A. B. is a postdoctoral fellow of the FWO-Vlaanderen.

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Authors and Affiliations

  1. Laboratory of Molecular and Cellular Therapy, Department of Immunology–Physiology and the Dendritic Cell Bank, Vrije Universiteit Brussel, Brussels, Belgium

    Daphné Benteyn, An M. T. Van Nuffel, Sofie Wilgenhof & Aude Bonehill

  2. Department of Medical Oncology, Universitair Ziekenhuis Brussel, Brussels, Belgium

    Sofie Wilgenhof

Authors
  1. Daphné Benteyn
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  2. An M. T. Van Nuffel
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  3. Sofie Wilgenhof
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  4. Aude Bonehill
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Editor information

Editors and Affiliations

  1. Morphogenesis, Inc., Tampa, Florida, USA

    Michael J.P. Lawman

  2. Morphogenesis, Inc., Tampa, Florida, USA

    Patricia D. Lawman

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Benteyn, D., Van Nuffel, A.M.T., Wilgenhof, S., Bonehill, A. (2014). Single-Step Antigen Loading and Maturation of Dendritic Cells Through mRNA Electroporation of a Tumor-Associated Antigen and a TriMix of Costimulatory Molecules. In: Lawman, M., Lawman, P. (eds) Cancer Vaccines. Methods in Molecular Biology, vol 1139. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-0345-0_1

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  • DOI: https://doi.org/10.1007/978-1-4939-0345-0_1

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-0344-3

  • Online ISBN: 978-1-4939-0345-0

  • eBook Packages: Springer Protocols

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