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In Vivo Electroporation Enhances the Immunogenicity of Hepatitis C Virus Nonstructural 3/4A DNA by Increased Local DNA Uptake, Protein Expression, Inflammation, and Infiltration of CD3+ T Cells

Gustaf Ahlén, Jonas Söderholm, Torunn Tjelle, Rune Kjeken, Lars Frelin, Urban Höglund, Pontus Blomberg, Michael Fons, Iacob Mathiesen and Matti Sällberg
J Immunol October 1, 2007, 179 (7) 4741-4753; DOI: https://doi.org/10.4049/jimmunol.179.7.4741
Gustaf Ahlén
*Division of Clinical Microbiology, F68, Karolinska Institutet at Karolinska University Hospital Huddinge, Stockholm, Sweden;
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Jonas Söderholm
*Division of Clinical Microbiology, F68, Karolinska Institutet at Karolinska University Hospital Huddinge, Stockholm, Sweden;
†Inovio Biomedical Corp., San Diego, CA 92121;
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Torunn Tjelle
‡Inovio AS, Gaustadalleen, Oslo, Norway;
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Rune Kjeken
†Inovio Biomedical Corp., San Diego, CA 92121;
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Lars Frelin
*Division of Clinical Microbiology, F68, Karolinska Institutet at Karolinska University Hospital Huddinge, Stockholm, Sweden;
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Urban Höglund
§Visionar AB, Uppsala, Sweden; and
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Pontus Blomberg
¶Vecura and Clinical Research Center, Karolinska University Hospital, Stockholm, Sweden
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Michael Fons
†Inovio Biomedical Corp., San Diego, CA 92121;
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Iacob Mathiesen
‡Inovio AS, Gaustadalleen, Oslo, Norway;
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Matti Sällberg
*Division of Clinical Microbiology, F68, Karolinska Institutet at Karolinska University Hospital Huddinge, Stockholm, Sweden;
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Abstract

The mechanisms by which in vivo electroporation (EP) improves the potency of i.m. DNA vaccination were characterized by using the hepatitis C virus nonstructural (NS) 3/4A gene. Following a standard i.m. injection of DNA with or without in vivo EP, plasmid levels peaked immediately at the site of injection and decreased by 4 logs the first week. In vivo EP did not promote plasmid persistence and, depending on the dose, the plasmid was cleared or almost cleared after 60 days. In vivo imaging and immunohistochemistry revealed that protein expression was restricted to the injection site despite the detection of significant levels of plasmid in adjacent muscle groups. In vivo EP increased and prolonged NS3/4A protein expression levels as well as an increased infiltration of CD3+ T cells at the injection site. These factors most likely additively contributed to the enhanced and broadened priming of NS3/4A-specific Abs, CD4+ T cells, CD8+ T cells, and γ-IFN production. The primed CD8+ responses were functional in vivo, resulting in elimination of hepatitis C virus NS3/4A-expressing liver cells in transiently transgenic mice. Collectively, the enhanced protein expression and inflammation at the injection site following in vivo EP contributed to the priming of in vivo functional immune responses. These localized effects most likely help to insure that the strength and duration of the responses are maintained when the vaccine is tested in larger animals, including rabbits and humans. Thus, the combined effects mediated by in vivo EP serves as a potent adjuvant for the NS3/4A-based DNA vaccine.

  • Received March 1, 2007.
  • Accepted July 18, 2007.
  • Copyright © 2007 by The American Association of Immunologists
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The Journal of Immunology: 179 (7)
The Journal of Immunology
Vol. 179, Issue 7
1 Oct 2007
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In Vivo Electroporation Enhances the Immunogenicity of Hepatitis C Virus Nonstructural 3/4A DNA by Increased Local DNA Uptake, Protein Expression, Inflammation, and Infiltration of CD3+ T Cells
Gustaf Ahlén, Jonas Söderholm, Torunn Tjelle, Rune Kjeken, Lars Frelin, Urban Höglund, Pontus Blomberg, Michael Fons, Iacob Mathiesen, Matti Sällberg
The Journal of Immunology October 1, 2007, 179 (7) 4741-4753; DOI: 10.4049/jimmunol.179.7.4741

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In Vivo Electroporation Enhances the Immunogenicity of Hepatitis C Virus Nonstructural 3/4A DNA by Increased Local DNA Uptake, Protein Expression, Inflammation, and Infiltration of CD3+ T Cells
Gustaf Ahlén, Jonas Söderholm, Torunn Tjelle, Rune Kjeken, Lars Frelin, Urban Höglund, Pontus Blomberg, Michael Fons, Iacob Mathiesen, Matti Sällberg
The Journal of Immunology October 1, 2007, 179 (7) 4741-4753; DOI: 10.4049/jimmunol.179.7.4741
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