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Dendritic cells transfected with interleukin-12 and tumor-associated antigen messenger RNA induce high avidity cytotoxic T cells

Gene Therapy volume 14, pages 366–375 (2007)Cite this article

Abstract

Dendritic cells (DC) transfected with messenger RNA (mRNA) encoding tumor-associated antigens (TAA) are able to induce potent tumor-specific T-cell responses directed to a broad spectrum of tumor-associated epitopes. The in vitro generation of DC possessing all the features crucial for the induction of type 1 immune responses, such as mature state, migratory potential and interleukin-12 (IL-12p70) production is complicated. Particularly migratory potential is inversely correlated with IL-12p70 production after maturation with prostaglandin E2 (PGE2), which is included in maturation cocktails currently used in most vaccination trials. Here, we show that transfection of PGE2 matured DC with a single mRNA strain encoding for ubiquitin followed by a TAA which was linked to IL-12 by a self-cleaving 2A sequence, produced biological active IL-12p70 and were able to present the transfected TAA up to 72 h after transfection. Furthermore, use of the anti-reverse cap analog for in vitro transcription of the IL-12 mRNA enabled constitutive IL-12p70 production for up to 5 days. These transfected mature DC migrated efficiently towards lymph node derived chemokines. DCs constitutively expressing IL-12p70, generate TAA-specific cytotoxic T cells with an high functional avidity, independent of CD4+ T-cell help.

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Acknowledgements

We thank A Stam for technical assistance, and the Maurits and Anna de Kock Foundation for financial support in the purchase of an HPLC. We thank Dr ML Kapsenberg for providing IL-12 antibodies and Dr S Hallez for providing the ubiquitin open-reading frame. We are grateful to Drs KJB Scholten and Dr MWJ Schreurs for fruitful discussions. This work was financially supported by Dutch Cancer Society (KWF) Grant VU2002-2627. VFIVT is a postdoctoral fellow of the fund for Scientific Research-Flanders (FWO-Vlaanderen).

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Authors and Affiliations

  1. Department of Pathology, Vrije Universiteit Medical Center, Amsterdam, The Netherlands

    H J Bontkes, D Kramer, J J Ruizendaal, E W M Kueter, C J L M Meijer & E Hooijberg

  2. Laboratory of Experimental Hematology, Faculty of Medicine, University of Antwerp, Antwerp University Hospital, Antwerp, Belgium

    V F I van Tendeloo

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Correspondence to H J Bontkes.

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Bontkes, H., Kramer, D., Ruizendaal, J. et al. Dendritic cells transfected with interleukin-12 and tumor-associated antigen messenger RNA induce high avidity cytotoxic T cells. Gene Ther 14, 366–375 (2007). https://doi.org/10.1038/sj.gt.3302874

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