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Transcriptional targeting of dendritic cells for gene therapy using the promoter of the cytoskeletal protein fascin

Gene Therapy volume 10, pages 1035–1040 (2003)Cite this article

Abstract

Strong cell-type-specific promoters are basic tools in gene therapy allowing for novel applications and focused strategies by transcriptionally targeting gene expression to selected cells. In immunotherapy, dendritic cells (DC) are of central importance, since they represent the principal inducers of immune responses. Here we describe isolation and use of the promoter of the murine actin-bundling protein fascin to target transcriptionally gene expression to cutaneous DC. Using the reporter gene enhanced green fluorescent protein (EGFP), we demonstrate that the fascin promoter mediates a strong antigen expression that is restricted to mature DC. DNA vaccination with antigen-encoding expression vectors under control of the fascin promoter using a gene gun resulted, consistently, in limited antigen expression by few directly transfected DC. Nevertheless, nearly as many antigen-specific CD8+ T cells directed against the encoded antigens EGFP and β-galactosidase, respectively, were induced as with expression constructs under control of the ubiquitously expressed CMV promoter. This result impressively underlines the pivotal role of directly transfected DC in DNA vaccination. Immunization using the fascin promoter induced markedly lower levels of antigen-specific antibodies following single or repeated immunization. Thus, our DC-targeted DNA vaccination approach induces qualitatively distinct, predominantly cellular immune responses and provides new opportunities for immunotherapy.

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Acknowledgements

This work was supported by the Deutsche Forschungsgemeinschaft, SFB 548 and SFB 432. We thank Dr Albert DeLeo (University of Pittsburgh School of Medicine, Pittsburgh, PA, USA) for providing the EGFP peptide, Dr Kenneth Rock (UMass Medical School, Worcester, MA, USA) for providing the cell line DC2.4, Drs Sem Saeland and Giorgio Trinchieri (both Schering-Plough Laboratory for Immunological Research, Dardilly, France) for providing the anti-langerin antibody 929F3 and Dr Akira Takashima (University of Texas Southwestern Medical Center, Dallas, TX, USA) for providing the cell line XS52. The expert technical assistance of Ms Katrin Krause & Ms Nadine Wiechmann is gratefully acknowledged.

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  1. J Steitz & T Tüting

    Present address: Department of Dermatology, University of Bonn, Sigmund-Freud-Str 25, 53105, Bonn, Germany

Authors and Affiliations

  1. Department of Dermatology, Clinical Research Unit Allergology, Johannes Gutenberg University, Mainz, Germany

    R Ross, S Sudowe, J Beisner, X-L Ross, I Ludwig-Portugall, J Steitz, T Tüting, J Knop & A B Reske-Kunz

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Ross, R., Sudowe, S., Beisner, J. et al. Transcriptional targeting of dendritic cells for gene therapy using the promoter of the cytoskeletal protein fascin. Gene Ther 10, 1035–1040 (2003). https://doi.org/10.1038/sj.gt.3301968

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