Hepatitis C virus (HCV) infections are a significant threat to human health. Currently, no vaccine exists against infection and/or the liver disease caused by HCV. Because a cell-culture system for the propagation of the virus is not available, modified live-virus or inactivated vaccine development has been impossible. The objective of the proposal presented here is to demonstrate that the equine herpesvirus type 1 (EHV-1) vaccine strain RacH can be developed into a vector that is able to efficiently deliver sequences encoding HCV structural proteins. We will also test, which route of application of the EHV-1 vector results in a significant humoral and cellular immune response against HCV. EHV-1 is a novel vaccine vector given its efficient entry into primary human peripheral blood mononuclear cells (PBMC), including professional antigen-presenting cells, its ability to infect at low particle per cell numbers, and the possibility to engineer extremely safe replication-deficient EHV-1 that can accommodate large pieces of foreign DNA. In addition, pre-existing immunity against EHV-1 in humans is absent.
Specific aim 1. To construct and characterize EHV-1 recombinants expressing HCV structural proteins C/E1 to p7. Recombinant viruses will be generated using the infectious bacterial artificial chromosome (BAG) clone of RacH (pRacH). We shall generate EHV-1 expressing the capsid to p7 (C-p7) or the El to p7 (El-p7) region of HCV. The HCV constructs will be inserted in two different genomic loci of the RacH genome, namely the unique-long or unique-short region, to exclude positional effects of foreign gene insertion. The same HCV structural gene region will also be inserted into an extremely safe replication-deficient RacH virus, which has a deletion in the sole and essential immediate-early gene and is unable to replicate or produce any EHV-1 proteins unless propagated in a complementing cell line.
Specific aim 2. To evaluate the immunogenic properties of recombinant EHV-1 expressing HCV proteins. The pRacH-based viruses expressing the HCV structural proteins will be tested for the induction of ELISA and neutralizing antibodies as well as cell-mediated cytotoxic (CTL) responses in mice. The tests will include ELISPOT and CTL assays in the context of a human class I allele using HLA-A2.1 transgenic mice.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Exploratory/Developmental Grants (R21)
Project #
5R21AI061412-02
Application #
7026477
Study Section
Special Emphasis Panel (ZRG1-VMD (01))
Program Officer
Koshy, Rajen
Project Start
2005-03-15
Project End
2007-02-28
Budget Start
2006-03-01
Budget End
2007-02-28
Support Year
2
Fiscal Year
2006
Total Cost
$175,000
Indirect Cost
Name
Cornell University
Department
Microbiology/Immun/Virology
Type
Schools of Veterinary Medicine
DUNS #
872612445
City
Ithaca
State
NY
Country
United States
Zip Code
14850
Goodwin, Thomas J; McCarthy, Maureen; Osterrieder, Nikolaus et al. (2013) Three-dimensional normal human neural progenitor tissue-like assemblies: a model of persistent varicella-zoster virus infection. PLoS Pathog 9:e1003512
Kaufer, Benedikt B; Smejkal, Benjamin; Osterrieder, Nikolaus (2010) The varicella-zoster virus ORFS/L (ORF0) gene is required for efficient viral replication and contains an element involved in DNA cleavage. J Virol 84:11661-9
Rosas, Cristina; Van de Walle, Gerlinde R; Metzger, Stephan M et al. (2008) Evaluation of a vectored equine herpesvirus type 1 (EHV-1) vaccine expressing H3 haemagglutinin in the protection of dogs against canine influenza. Vaccine 26:2335-43
Rosas, Cristina T; Paessler, Slobodan; Ni, Haolin et al. (2008) Protection of mice by equine herpesvirus type 1 based experimental vaccine against lethal Venezuelan equine encephalitis virus infection in the absence of neutralizing antibodies. Am J Trop Med Hyg 78:83-92
Rosas, Cristina T; Tischer, B Karsten; Perkins, Gillian A et al. (2007) Live-attenuated recombinant equine herpesvirus type 1 (EHV-1) induces a neutralizing antibody response against West Nile virus (WNV). Virus Res 125:69-78
Tischer, B Karsten; Kaufer, Benedikt B; Sommer, Marvin et al. (2007) A self-excisable infectious bacterial artificial chromosome clone of varicella-zoster virus allows analysis of the essential tegument protein encoded by ORF9. J Virol 81:13200-8
Rosas, Cristina T; Konig, Patricia; Beer, Martin et al. (2007) Evaluation of the vaccine potential of an equine herpesvirus type 1 vector expressing bovine viral diarrhea virus structural proteins. J Gen Virol 88:748-57