title: [DNA immunization with a plasmid containing gene of hepatitis C virus protein 5A (NS5A) induces the effective cellular immune response].
abstract: In spite of extensive research, no effective vaccine against hepatitis C virus (HCV) has been developed so far. DNA immunization is a potent technique of vaccine design strongly promoting the cellular arm of immune response. The genes encoding nonstructural HCV proteins (NS2-NS5B) are promising candidates for vaccine development. NS5A is a protein involved in viral pathogenesis, in the induction of immune response, and probably in viral resistance to interferon treatment. The objective of this study was to construct a DNA vaccine encoding NS5A protein and evaluate its immunogenicity. A plasmid encoding a full-size NS5A protein was produced using the pcDNA3.1 (+) vector for eukaryotic expression system. The expression of the NS5A gene was confirmed by immunoperoxidase staining of the transfected eukaryotic cells with anti-NS5A monoclonal antibodies. Triple immunization of mice with the plasmid vaccine induced a pronounced cellular immune response against abroad spectrum of NSSA epitopes as assessed by T-cell proliferation andsecretion of antiviral cytokines IFN-gamma and IL-2. In in vitro T-cell stimulation experiments, NS5A-derived antigens were modeled by synthetic peptides, recombinant proteins of various genotypes, and phages carrying exposed NS5A peptides. A novel immunomodulator Immunomax showed high adjuvant activity in DNA immunization. The data obtained indicate that the suggested DNA construct has a strong potential in the development of the gene vaccines against hepatitis C.
authors.forename: O V
authors.initials: OV
authors.lastname: Masalova
authors.forename: E I
authors.initials: EI
authors.lastname: Lesnova
authors.forename: V V
authors.initials: VV
authors.lastname: Grabovetskiĭ
authors.forename: O A
authors.initials: OA
authors.lastname: Smirnova
authors.forename: T I
authors.initials: TI
authors.lastname: Ulanova
authors.forename: A N
authors.initials: AN
authors.lastname: Burkov
authors.forename: A V
authors.initials: AV
authors.lastname: Ivanov
authors.forename: A D
authors.initials: AD
authors.lastname: Zaberezhnyĭ
authors.forename: R I
authors.initials: RI
authors.lastname: Ataullakhanov
authors.forename: A A
authors.initials: AA
authors.lastname: Kushch
mesh_terms: D000818:Animals; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D002522:Chlorocebus aethiops; D005260:Female; D016174:Hepacivirus / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D006526:Hepatitis C / Q000235:genetics / Q000276:immunology* / Q000378:metabolism / Q000517:prevention & control; D006801:Humans; D007111:Immunity, Cellular; D007371:Interferon-gamma / Q000096:biosynthesis / Q000276:immunology; D007376:Interleukin-2 / Q000096:biosynthesis / Q000276:immunology; D051379:Mice; D010957:Plasmids / Q000235:genetics / Q000276:immunology / Q000378:metabolism / Q000494:pharmacology*; D013601:T-Lymphocytes / Q000276:immunology* / Q000378:metabolism; D019444:Vaccines, DNA / Q000235:genetics / Q000276:immunology / Q000378:metabolism / Q000494:pharmacology*; D014709:Vero Cells; D014761:Viral Hepatitis Vaccines / Q000235:genetics / Q000276:immunology / Q000378:metabolism / Q000494:pharmacology*; D017361:Viral Nonstructural Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*; D012324:RNA-Dependent RNA Polymerase
chemical_list: D007371:Interferon-gamma; D007376:Interleukin-2; D019444:Vaccines, DNA; D014761:Viral Hepatitis Vaccines; D017361:Viral Nonstructural Proteins; C087036:NS-5 protein, hepatitis C virus; D012324:RNA-Dependent RNA Polymerase