title: The structural HCV genes delivered by MPG cell penetrating peptide are directed to enhance immune responses in mice model.
abstract: One of the significant problems in vaccination projects is the lack of an effective vaccine against hepatitis C virus (HCV). The goal of the current study is to evaluate and compare two DNA constructs encoding HCV core and coreE1E2 genes alone or complexed with MPG peptide as a delivery system for stimulation of antibody responses and IFN-γ secretion in Balb/c mice model. Indeed, MPG cell penetrating peptide was used to improve DNA immunization in mice. Our results demonstrated that MPG forms stable non-covalent nanoparticles with pcDNA-core and pcDNA-coreE1E2 at an N/P ratio of 10:1. The in vitro transfection efficiency of core or coreE1E2 DNA using MPG and TurboFect delivery systems was confirmed by western blot analysis. The results indicated the expression of the full-length core (∼21 kDa), and coreE1E2 (∼83 kDa) proteins using an anti-His monoclonal antibody. In addition, the expression of HCV core and coreE1E2 proteins was performed in bacteria and the purified recombinant proteins were injected to mice with Montanide 720 adjuvant. Our data showed that the immunized mice with HCV core and coreE1E2 proteins generated the mixture of sera IgG1 and IgG2a isotypes considerably higher than other groups. Furthermore, DNA constructs encoding core and coreE1E2 complexed with MPG could significantly induce IFN-γ secretion in lower concentrations than the naked core and coreE1E2 DNAs. Taken together, the DNA formulations as well as protein regimens used in this study triggered high-level IFN-γ production in mice, an important feature for the development of Th1 immune responses.
authors.affiliation: a Department of Hepatitis and AIDS , Pasteur Institute of Iran , Tehran , Iran.|b Department of Biotechnology , Faculty of Advanced Sciences and Technology, Pharmaceutical Sciences Branch, Islamic Azad University , Tehran , Iran , and.
authors.forename: Saloume
authors.initials: S
authors.lastname: Mehrlatifan
authors.affiliation: c Department of Biology , Science and Research Branch, Islamic Azad University , Tehran , Iran.
authors.forename: Seyyedeh Masumeh
authors.initials: SM
authors.lastname: Mirnurollahi
authors.affiliation: a Department of Hepatitis and AIDS , Pasteur Institute of Iran , Tehran , Iran.
authors.forename: Fatemeh
authors.initials: F
authors.lastname: Motevalli
authors.affiliation: a Department of Hepatitis and AIDS , Pasteur Institute of Iran , Tehran , Iran.
authors.forename: Pooneh
authors.initials: P
authors.lastname: Rahimi
authors.affiliation: a Department of Hepatitis and AIDS , Pasteur Institute of Iran , Tehran , Iran.
authors.forename: Sepehr
authors.initials: S
authors.lastname: Soleymani
authors.affiliation: a Department of Hepatitis and AIDS , Pasteur Institute of Iran , Tehran , Iran.
authors.forename: Azam
authors.initials: A
authors.lastname: Bolhassani
mesh_terms: D000276:Adjuvants, Immunologic; D000818:Animals; D002460:Cell Line; D057846:Cell-Penetrating Peptides / Q000235:genetics* / Q000276:immunology*; D004268:DNA-Binding Proteins / Q000235:genetics* / Q000276:immunology*; D005260:Female; D005814:Genes, Viral / Q000235:genetics* / Q000276:immunology*; D057809:HEK293 Cells; D016174:Hepacivirus / Q000235:genetics / Q000276:immunology*; D006526:Hepatitis C / Q000276:immunology; D006801:Humans; D007111:Immunity, Cellular / Q000276:immunology; D007114:Immunization / Q000379:methods; D007074:Immunoglobulin G / Q000276:immunology; D007371:Interferon-gamma / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D014162:Transfection / Q000379:methods; D019444:Vaccines, DNA / Q000235:genetics / Q000276:immunology
chemical_list: D000276:Adjuvants, Immunologic; D057846:Cell-Penetrating Peptides; D004268:DNA-Binding Proteins; D007074:Immunoglobulin G; C475074:MPG peptide; D019444:Vaccines, DNA; D007371:Interferon-gamma
keywords: Cell penetrating cell; DNA immunization; HCV; MPG; delivery system; protein immunization