Interleukin-12 as a genetic adjuvant enhances hepatitis C virus NS3 DNA vaccine immunogenicity
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Hepatitis C virus (HCV) chronic infection is a worldwide health problem, and numerous efforts have been invested to develop novel vaccines. An efficient vaccine requires broad immune response induction against viral proteins. To achieve this goal, we constructed a DNA vaccine expressing nonstructural 3 (NS3) gene (pcDNA3.1-HCV-NS3) and assessed the immune response in C57BL/6 mice. In this study, the NS3 gene was amplified with a nested-reverse transcriptase-polymerase chain reaction (RT-PCR) method using sera of HCV-infected patients with genotype 1a. The resulting NS3 gene was subcloned into a pcDNA3.1 eukaryotic expression vector, and gene expression was detected by western blot. The resultant DNA vaccine was co-administered with interleukin-12 (IL-12) as an adjuvant to female C57BL/6 mice. After the final immunizations, lymphocyte proliferation, cytotoxicity, and cytokine levels were assessed to measure immune responses. Our data suggest that co-administration of HCV NS3 DNA vaccine with IL-12 induces production of significant levels of both IL-4 and interferon (IFN)-γ (p<0.05). Cytotoxicity and lymphocyte proliferation responses of vaccinated mice were significantly increased compared to control (p<0.05). Collectively, our results demonstrated that co-administration of HCV NS3 and IL-12 displayed strong immunogenicity in a murine model.
KeywordsHepatitis C virus (HCV) NS3 Interleukin-12 DNA vaccine
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- Ahlen G, Söderholm J, Tjelle T, Kjeken R, Frelin L, Höglund U, Blomberg P, Fons M, Mathiesen I, and Sällberg M. 2007. 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. J Immunol, 179(7): 4741–4753.PubMedGoogle Scholar
- Arribillaga L, de Cerio A L, Sarobe P, Casares N, Gorraiz M, Vales A, Bruna-Romero O, Borrás-Cuesta F, Paranhos-Baccala G, Prieto J, Ruiz J, and Lasarte J J. 2002. Vaccination with an adenoviral vector encoding hepatitis C virus (HCV) NS3 protein protects against infection with HCV recombinant vaccinia virus. Vaccine, 21: 202–210.PubMedCrossRefGoogle Scholar
- Bocher W O, Dekel B, Schwerin W, Geissler M, Hoffmann S, Rohwer A, Arditti F, Cooper A, Bernhard H, Berrebi A, Rose-John S, Shaul Y, Galle P R, F Löhr H, and Reisner Y. 2001. Induction of strong hepatitis B virus (HBV) specific T helper cell and cytotoxic T lymphocyte responses by therapeutic vaccination in the trimera mouse model of chronic HBV infection. Eur J Immunol, 31: 2071–2079.PubMedCrossRefGoogle Scholar
- Encke J, zuPutlitz J, Geissler M, and R Wands J. 1998. Genetic Immunization generates cellular and Humoral Immune Responses Against the Nonstructural Proteins of the Hepatitis C Virus in a Murine Model. J Immunol, 1(61): 4917–4923.Google Scholar
- Ghaemi A, Soleimanjahi H, Gill P, Zuhair H, Razeghi M, Jahromi S, and Roohvand F. 2010. Recombinant λ-phage nanobioparticles for tumor therapy in mice models. J Genet Vaccines Ther, 8(3): 1–7.Google Scholar
- Gherardi M M, Ramirez J C, and Esteban M. 2000. Interleukin-12 (IL-12) Enhancement of the Cellular Immune Response against Human Immunodeficiency Virus Type 1 Env Antigen in a DNA Prime/Vaccinia Virus Boost VaccineRegimen Is Time and Dose Dependent: Suppressive Effects of IL-12 Boost Are Mediated by Nitric Oxide. J Virol, 74: 6278–6286.PubMedCrossRefGoogle Scholar
- Jiao X, Wang R Y, Feng Z, Hu G, Alter H J, and W-K Shih J. DNA immunization encoding the secreted nonstructural protein 3 (NS3) of hepatitis C virus and enhancing the Th1 type immune response. 2004. J Viral Hepatol, 11(1):18–26.Google Scholar
- Sin J I, Kim J J, Arnold R L, Shroff K E, McCallus D, Pachuk C, McElhiney S P, Wolf M W, Pompa-de Bruin S J, Higgins T J, Ciccarelli R B, and Weiner D B. 1999. IL-12 Gene as a DNA Vaccine Adjuvant in a Herpes Mouse Model: IL-12 Enhances Th1-Type CD 4+ T Cell-Mediated Protective Immunity against Herpes Simplex Virus-2 Challenge. J Immunol, 162: 2912–2921.PubMedGoogle Scholar
- Weiner A J, Paliard X, Selby M J, Medina-Selby A, Coit D, Nguyen S, Kansopon Joe, L Arian Ch, Ng Ph, Tucker Jeffery, Lee Ch-T, K Polakos N, Han J, Wong Sh, Lu H-H, Rosenberg S, M Brasky K, Chien D, Kuo G, and Houghton M. 2001. Intrahepatic genetic inoculation of hepatitis C virus RNA confers cross-protective immunity. J Virol, 75(15): 7142–7148.PubMedCrossRefGoogle Scholar
- Yang S H, Lee C G, Park S H, Im S J, Kim Y M, Son J M, Wang J S, Yoon S K, Song M K, Ambrozaitis A, Kharchenko N, Yun Y D, Kim C M, Kim C Y, Lee S H, Kim B M, Kim W B, and Sung Y C. 2006. Correlation of antiviral Tcell responses with suppression of viral rebound in chronic hepatitis B carriers: a proof-of-concept study. J Gene Ther, 13(14): 1110–1117.CrossRefGoogle Scholar