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Low dose and gene gun immunization with a hepatitis C virus nonstructural (NS) 3 DNA-based vaccine containing NS4A inhibit NS3/4A-expressing tumors in vivo

Gene Therapy volume 10, pages 686–699 (2003)Cite this article

Abstract

The hepatitis C virus (HCV) protease and helicase encompasses the nonstructural (NS) 3 protein and the cofactor NS4A, which targets the NS3/4A-complex to intracellular membranes. We here evaluate the importance of NS4A in NS3-based genetic immunogens. A full-length genotype 1 NS3/4A gene was cloned into a eucaryotic expression vector in the form of NS3/4A and NS3 alone. Transient transfections revealed that the inclusion of NS4A increased the expression levels of NS3. Subsequently, immunization with the NS3/4A gene primed 10- to 100-fold higher levels of NS3-specific antibodies as compared to immunization with the NS3 gene. Humoral responses primed by the NS3/4A gene had a higher IgG2a/IgG1 ratio (>20) as compared to the NS3 gene (3.0), suggesting a T helper 1-skewed response. Low dose i.m. (10 μg) immunization with the NS3/4A gene inhibited the growth of NS3/4A-expressing tumor cells in vivo, whereas the NS3 gene alone or NS3 protein did not. We then evaluated the efficiency of the NS3/4A gene administered by the gene gun, at the same doses used for humans, in priming cytotoxic T lymphocyte (CTL) responses. Three to four 4 μg doses of the NS3/4A gene primed CTL at a precursor frequency of 2–4%, which inhibited the growth of NS3/4A-expressing tumor cells in vivo. Thus, NS4A enhances the expression levels and immunogenicity of NS3, and an NS3/4A gene delivered transdermally could be a therapeutic vaccine candidate.

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Acknowledgements

This study was supported by grant no. K2000-06X-12617-03A and K2002-16X-09494-12B from the Swedish Research Council, and by grant no. QLK2-1999-00588 from the European Community. BR was supported by the Wallenberg Consortia North for Functional Genomics.

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Authors and Affiliations

  1. Division of Clinical Virology, F 68, Karolinska Institutet at Huddinge University Hospital, Stockholm, Sweden

    L Frelin, M Alheim, A Chen, J Söderholm & M Sällberg

  2. Division of Pathology and Clinical Research Centre, Karolinska Institutet at Huddinge University Hospital, Stockholm, Sweden

    B Rozell

  3. Swedish Institute for Infectious Disease Control, Stockholm, Sweden

    C Barnfield & P Liljeström

  4. Microbiology and Tumourbiology Centre, Karolinska Institutet, Stockholm, Sweden

    P Liljeström

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Frelin, L., Alheim, M., Chen, A. et al. Low dose and gene gun immunization with a hepatitis C virus nonstructural (NS) 3 DNA-based vaccine containing NS4A inhibit NS3/4A-expressing tumors in vivo. Gene Ther 10, 686–699 (2003). https://doi.org/10.1038/sj.gt.3301933

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