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Liver-specific expression of interferon γ following adenoviral gene transfer controls hepatitis B virus replication in mice

Gene Therapy volume 12, pages 668–677 (2005)Cite this article

Abstract

Interferons control viral replication and the growth of some malignant tumors. Since systemic application may cause severe adverse effects, tissue-specific expression is an attractive alternative. Liver-directed interferon gene therapy offers promising applications such as chronic viral hepatitis B or C or hepatocellular carcinoma and thus needs testing in vivo in suitable animal models. We therefore used the Tet-On system to regulate gene expression in adenoviral vectors, and studied the effect of liver-specific and regulated interferon γ expression in a mouse model of chronic hepatitis B virus (HBV) infection. In a first generation adenoviral vector, genes encoding for firefly luciferase and interferons α, β or γ, respectively, were coexpressed under control of the bidirectional tetracycline-regulated promoter Ptetbi. Liver-specific promoters driving expression of the reverse tetracycline controlled transactivator ensured local expression in the livers of HBV transgenic mice. Following gene transfer, we demonstrated low background, tight regulation and a 1000-fold induction of gene expression by doxycycline. Both genes within the bidirectional transcription unit were expressed simultaneously, and in a liver-specific fashion in cell culture and in living mice. Doxycycline-dependent interferon γ expression effectively controlled HBV replication in mice, but did not eliminate HBV transcripts. This system will help to study the effects of local cytokine expression in mouse disease models in detail.

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Acknowledgements

We thank the animal care unit of the Center of Molecular Biology Heidelberg for their highly motivated animal care, R Eilers for technical assistance, and U Baron for help with setting up the Tet-system. We thank H-G Kräusslich and H Schaller for critical discussion of the data and for continuous support. This work was supported by grant PR 618/2 from the Deutsche Forschungs-gemeinschaft to UP and by a grant from the Dr Mildred Scheel Stiftung to PS.

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Authors and Affiliations

  1. Department of Virology, University of Heidelberg, Heidelberg, Germany

    J Dumortier, H Oberwinkler & U Protzer

  2. Center of Molecular Biology, University of Heidelberg, Heidelberg, Germany

    K Schönig, R Löw & H Bujard

  3. Central Institute of Mental Health, Mannheim, Germany

    K Schönig

  4. Molecular Infectiology at the Center for Molecular Medicine Cologne, Institute of Medical Microbiology, University of Cologne, Cologne, Germany

    H Oberwinkler & U Protzer

  5. Institute of Immunology, University of Heidelberg, Heidelberg, Germany

    T Giese

  6. Institute of Pathology, University of Cologne, Cologne, Germany

    P Schirmacher

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Dumortier, J., Schönig, K., Oberwinkler, H. et al. Liver-specific expression of interferon γ following adenoviral gene transfer controls hepatitis B virus replication in mice. Gene Ther 12, 668–677 (2005). https://doi.org/10.1038/sj.gt.3302449

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