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Regulated and constitutive expression of anti-inflammatory cytokines by nontransforming herpesvirus saimiri vectors

Gene Therapy volume 12, pages 395–406 (2005)Cite this article

Abstract

Herpesviral saimiri-(HVS) mediated expression of bovine growth hormone was one of the first applications of an episomal viral vector for gene therapy. Meanwhile, the long-term persistence of HVS vectors has been confirmed in a broad spectrum of infectable target cells in vitro and in vivo. Regulated gene expression is useful for many applications of gene therapy. Therefore, we inserted the Mifepristone–antiprogestin-inducible expression system (GeneSwitch™) into HVS viral vectors to regulate the combined expression of anti-inflammatory cytokines, IL-10 and IL-1RA. Constitutive CMV-promoter/enhancer-driven and Mifepristone-inducible cytokine expression was compared in the viral context in transduced primary human fibroblasts and rheumatoid arthritis (RA) fibroblast-like cells (RASF). Long-term persistence of vector genomes was shown for both construct types. Constitutive expression was efficient and more rapid in onset than in the inducible system, in which the selective induction of interleukin expression along with low background levels was obtained by Mifepristone concentrations that were more than 1000-fold below those required for endogenous Progesterone antagonism. Furthermore, transgene expression corresponded to vector doses. Global patterns of cytokine secretion were not significantly changed due to viral transduction, indicating a rather inert behavior of the viral vector itself. In an attempt to emulate the inflammatory cytokine-enriched environment in rheumatoid arthritic joints, the function of the vectors could be demonstrated in vitro by the successful blockade of IL-1β-stimulated matrix-metalloproteinase (MMP)-3 expression from RASF cells. Evaluation of this system in future studies, in suitable long-term SCID models of RA or in non-human primate models, will exploit the possible in vivo benefits of nontransforming HVS vectors in gene therapy.

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Acknowledgements

We are grateful to Alexandra Schäfer and Elke Heck for critical reading of the manuscript. This project was supported by the Interdisciplinary Center for Clinical Research (IZKF: Genesis, Diagnostics and Therapy of Inflammation Processes) at the University of Erlangen-Nuremberg, the Deutsche Forschungsgemeinschaft (Sonderforschungsbereich 466), the Federal Ministry of Education and Research (BMBF) and the Wilhelm Sander-Stiftung.

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Authors and Affiliations

  1. Institut für Klinische und Molekulare Virologie, Universität Erlangen-Nürnberg, Erlangen, Germany

    C Wieser, D Stumpf, C Grillhösl, D Lengenfelder, B Fleckenstein & A Ensser

  2. Zentrum für Experimentelle Rheumatologie, Universität Zürich, Switzerland

    S Gay

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  1. C Wieser
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Wieser, C., Stumpf, D., Grillhösl, C. et al. Regulated and constitutive expression of anti-inflammatory cytokines by nontransforming herpesvirus saimiri vectors. Gene Ther 12, 395–406 (2005). https://doi.org/10.1038/sj.gt.3302424

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