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Journal of Virology, January 2003, p. 382-390, Vol. 77, No. 1
0022-538X/03/$08.00+0     DOI: 10.1128/JVI.77.1.382-390.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

DNA Immunization with Hepatitis C Virus (HCV) Polycistronic Genes or Immunization by HCV DNA Priming-Recombinant Canarypox Virus Boosting Induces Immune Responses and Protection from Recombinant HCV-Vaccinia Virus Infection in HLA-A2.1-Transgenic Mice

Preeti Pancholi,1,{dagger} Marion Perkus,2 Nancy Tricoche,1 Qingyan Liu,1 and Alfred M. Prince1*

Laboratory of Virology, Lindsley F. Kimball Research Institute of the New York Blood Center, New York,1 Virogenetics, Troy, New York2

Received 28 May 2002/ Accepted 24 September 2002

We studied immune responses to hepatitis C virus (HCV) genes delivered as DNA encoding the entire HCV protein coding genome in two polycistronic plasmids encoding HCV capsid-E1-E2-NS2-NS3 and HCV NS3-NS4-NS5 in HLA-A2.1-transgenic mice. Immune responses to HCV DNA prime and recombinant canarypox virus boost were also studied with the above constructs. At 8 weeks after a canarypox virus boost, the DNA prime/canarypox virus boosting regimen induced potent cellular immune responses to HCV structural and nonstructural proteins on target cells expressing the HLA-A2.1 allele. High frequencies of gamma interferon-secreting cells, as detected by enzyme-linked immunospot assay, were obtained in response to several endogenously expressed HCV proteins. We also observed cytotoxic-T-lymphocyte reactivity in response to endogenously expressed HCV proteins in fresh spleen cells without in vitro expansion. Upon challenge with a recombinant vaccinia virus expressing HCV proteins at 2 months postimmunization, the HCV DNA prime/canarypox virus-immunized mice showed a complete reduction in vaccinia virus titers compared to HCV DNA prime/boost- and mock-immunized controls. Immune responses were still detectable 4 months after canarypox virus boost in immunized mice. Interestingly, at 10 months postimmunization (8 months after canarypox virus boost), the protection in HCV DNA prime/boost-immunized mice against recombinant HCV-vaccinia virus challenge was higher than that observed in HCV DNA prime/canarypox virus boost-immunized mice.

* Corresponding author. Mailing address: Laboratory of Virology, Lindsley F. Kimball Research Institute, New York Blood Center, 310 E. 67th St., New York, NY 10021. Phone: (212) 570-3279. Fax: (212) 570-3180. E-mail: aprince{at}nybc.org.

{dagger} Present address: Columbia-Presbyterian Medical Center, New York, NY 10032.

Journal of Virology, January 2003, p. 382-390, Vol. 77, No. 1
0022-538X/03/$08.00+0     DOI: 10.1128/JVI.77.1.382-390.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.





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Copyright © 2003 by the American Society for Microbiology. All rights reserved.