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Intradermal delivery of DNA encoding HCV NS3 and perforin elicits robust cell-mediated immunity in mice and pigs

Gene Therapy volume 23, pages 26–37 (2016)Cite this article

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Abstract

Currently, no vaccine is available against hepatitis C virus (HCV), and although DNA vaccines have considerable potential, this has not been realised. Previously, the efficacy of DNA vaccines for human immunodeficiency virus (HIV) and HCV was shown to be enhanced by including the gene for a cytolytic protein, viz. perforin. In this study, we examined the mechanism of cell death by this bicistronic DNA vaccine, which encoded the HCV non-structural protein 3 (NS3) under the control of the CMV promoter and perforin is controlled by the SV40 promoter. Compared with a canonical DNA vaccine and a bicistronic DNA vaccine encoding NS3 and the proapoptotic gene NSP4, the perforin-containing vaccine elicited enhanced cell-mediated immune responses against the NS3 protein in vaccinated mice and pigs, as determined by ELISpot and intracellular cytokine staining, whereas a mouse challenge model suggested that the immunity was CD8+ T-cell-dependent. The results of the study showed that the inclusion of perforin in the DNA vaccine altered the fate of NS3-positive cells from apoptosis to necrosis, and this resulted in more robust immune responses in mice and pigs, the latter of which represents an accepted large animal model in which to test vaccine efficacy.

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Acknowledgements

We thank Dr Paul Radspinner (FluGen Inc., Madison, WI, USA) who provided the microneedle device and Renee Herber for training to use the device. We also thank Dr John Taylor (University of Auckland, New Zealand) and Dr Barbara Coulson (University of Melbourne, Australia) for the gift of the NSP4 construct. We acknowledge the kind gift of the HCV peptide pools, which were obtained through the AIDS Reagent and Reference Reagent Program, Division of AIDS, NIAID, National Institutes of Health, USA. We also thank Dr Stuart Howell for statistical advice. This research was supported by grants APP1026293, 543139 and 543143 from the National Health and Medical Research Council (NHMRC) of Australia, grant BF040005 from the Australian-Indian biotechnology fund and a grant from the Hospital Research Foundation (THRF). DW is a Research Fellow supported by THRF.

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Author notes

  1. W Yu

    Present address: 4Current address: Experimental Therapeutics Laboratory, University of South Australia, Adelaide, SA, Australia.,

  2. T Garrod

    Present address: 5Current address: Royal Australasian College of Surgeons, North Adelaide, SA, Australia.,

  3. B Grubor-Bauk and W Yu: These authors contributed equally to this work.

Authors and Affiliations

  1. Virology Laboratory, Discipline of Surgery, University of Adelaide and Basil Hetzel Institute for Translational Medicine, Adelaide, SA, Australia

    B Grubor-Bauk, W Yu, D Wijesundara, J Gummow, T Garrod & E J Gowans

  2. Killer Cell Biology Laboratory, Cancer Immunology Research, Peter MacCallum Cancer Centre, Melbourne, VIC, Australia

    A J Brennan & I Voskoboinik

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  1. B Grubor-Bauk
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Grubor-Bauk, B., Yu, W., Wijesundara, D. et al. Intradermal delivery of DNA encoding HCV NS3 and perforin elicits robust cell-mediated immunity in mice and pigs. Gene Ther 23, 26–37 (2016). https://doi.org/10.1038/gt.2015.86

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