title: Evaluating replication-defective vesicular stomatitis virus as a vaccine vehicle.
abstract: We have generated replication-competent (VSV-C/E1/E2) and nonpropagating (VSVDeltaG-C/E1/E2) vesicular stomatitis virus (VSV) contiguously expressing the structural proteins of hepatitis C virus (HCV; core [C] and glycoproteins E1 and E2) and report on their immunogenicity in murine models. VSV-C/E1/E2 and VSVDeltaG-C/E1/E2 expressed high levels of HCV C, E1, and E2, which were authentically posttranslationally processed. Both VSV-expressed HCV E1-E2 glycoproteins were found to form noncovalently linked heterodimers and appeared to be correctly folded, as confirmed by coimmunoprecipitation analysis using conformationally sensitive anti-HCV-E2 monoclonal antibodies (MAbs). Intravenous or intraperitoneal immunization of BALB/c mice with VSV-C/E1/E2 or VSVDeltaG-C/E1/E2 resulted in significant and surprisingly comparable HCV core or E2 antibody responses compared to those of control mice. In addition, both virus types generated HCV C-, E1-, or E2-specific gamma interferon (IFN-gamma)-producing CD8(+) T cells, as determined by enzyme-linked immunospot (ELISPOT) analysis. Mice immunized with VSVDeltaG-C/E1/E2 were also protected against the formation of tumors expressing HCV E2 (CT26-hghE2t) and exhibited CT26-hghE2t-specific IFN-gamma-producing and E2-specific CD8(+) T-cell activity. Finally, recombinant vaccinia virus (vvHCV.S) expressing the HCV structural proteins replicated at significantly lower levels when inoculated into mice immunized with VSV-C/E1/E2 or VSVDeltaG-C/E1/E2, but not with control viruses. Our data therefore illustrate that potentially safer replication-defective VSV can be successfully engineered to express high levels of antigenically authentic HCV glycoproteins. In addition, this strategy may therefore serve in effective vaccine and immunotherapy-based approaches to the treatment of HCV-related disease.
authors.affiliation: Department of Microbiology and Immunology, University of Miami School of Medicine, FL 33136, USA.
authors.forename: Ayaz M
authors.initials: AM
authors.lastname: Majid
authors.forename: Heather
authors.initials: H
authors.lastname: Ezelle
authors.forename: Sangeeta
authors.initials: S
authors.lastname: Shah
authors.forename: Glen N
authors.initials: GN
authors.lastname: Barber
mesh_terms: D000818:Animals; D000917:Antibody Formation / Q000276:immunology; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology; D006224:Cricetinae; D004195:Disease Models, Animal; D005069:Evaluation Studies as Topic; D005260:Female; D005822:Genetic Vectors; D006367:HeLa Cells; D017325:Hepatitis B Vaccines / Q000235:genetics / Q000276:immunology* / Q000494:pharmacology; D006526:Hepatitis C / Q000276:immunology* / Q000517:prevention & control; D018937:Hepatitis C Antibodies / Q000276:immunology; D006801:Humans; D007114:Immunization; D007371:Interferon-gamma / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D011487:Protein Conformation; D011499:Protein Processing, Post-Translational / Q000235:genetics / Q000276:immunology; D014616:Vaccinia virus / Q000235:genetics / Q000276:immunology; D014721:Vesicular stomatitis Indiana virus / Q000235:genetics / Q000276:immunology*; D014759:Viral Envelope Proteins / Q000276:immunology*; D014779:Virus Replication
chemical_list: D017325:Hepatitis B Vaccines; D018937:Hepatitis C Antibodies; D014759:Viral Envelope Proteins; D007371:Interferon-gamma