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Cotransfection of dendritic cells with RNA coding for HER-2/neu and 4-1BBL increases the induction of tumor antigen specific cytotoxic T lymphocytes

Cancer Gene Therapy volume 12, pages 749–756 (2005)Cite this article

Abstract

Ribonucleic acid (RNA) transfection of dendritic cells (DCs) was shown to be highly efficient in eliciting CD8+ and CD4+ T-cell responses. We analyzed whether electroporation of DCs with RNA coding for a tumor-associated antigen (TAA) would elicit antigen-specific effector cytotoxic T lymphocyte (CTL) responses and whether these responses could be modulated by cotransfection with a second specific synthetic RNA. Therefore in vitro generated human monocyte-derived DCs were electroporated with in vitro transcribed RNA (in vitro transcript, IVT) encoding the TAA HER-2/neu. Additionally, these cells were cotransfected with IVT coding for human 4-1BBL. Transfection of DCs with 4-1BBL-IVT did not alter their typical phenotype. However, it increased the expression of the costimulatory molecules CD80 and CD40. Coadministration of HER-2/neu- and 4-1BBL-IVT resulted in an increased specific lysis of target cells by the in vitro induced CTL lines, indicating that 4-1BBL enhances their ability to elicit primary CTL responses. Interestingly, transfection of DCs with 4-1BBL-IVT did not augment their capacity to stimulate allogeneic lymphocyte responses. The here established approach of cotransfection of DCs with tumor-RNA and a second specific IVT could improve and optimize the in vitro manipulation of DCs for the induction of antigen-specific CTL responses.

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Acknowledgements

We thank Bruni Schuster, Sylvia Stephan, and Regina Heselmaier for excellent technical assistance. This work was supported by Deutsche Forschungsgemeinschaft (SFB 510, Project B2) and University of Tübingen (fortüne-project, F.1282620.1).

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  1. Frank Grünebach and Katrin Kayser: Both these authors contributed equally to this study.

Authors and Affiliations

  1. Department of Internal Medicine II, Hematology, Oncology and Immunology, University of Tübingen, Tübingen, Germany

    Frank Grünebach, Katrin Kayser, Markus M Weck, Martin R Müller, Silke Appel & Peter Brossart

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  1. Frank Grünebach
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  2. Katrin Kayser
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  3. Markus M Weck
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Correspondence to Peter Brossart.

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Grünebach, F., Kayser, K., Weck, M. et al. Cotransfection of dendritic cells with RNA coding for HER-2/neu and 4-1BBL increases the induction of tumor antigen specific cytotoxic T lymphocytes. Cancer Gene Ther 12, 749–756 (2005). https://doi.org/10.1038/sj.cgt.7700842

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