Title

Analysis of vesicular stomatitis virus expressing hepatitis C virus structural proteins as a potential vaccine

Date of Award

2005

Availability

Article

Degree Name

Doctor of Philosophy (Ph.D.)

Department

Microbiology and Immunology

First Committee Member

Glen N. Barber, Committee Chair

Abstract

Hepatitis C virus (HCV) currently infects over 170 million people worldwide and is a leading cause of liver disease, cirrhosis, hepatocellular carcinoma, and liver failure. Despite advances in standard therapy, only 40--50% of patients treated are able to resolve the infection, making the development of an effective vaccine a worldwide priority. In this study, we have generated a recombinant vesicular stomatitis virus (VSV) that expresses the polypeptide encoding the HCV structural proteins (VSV-HCV-C/E1/E2). This contiguous peptide is cleaved by signal peptidases into the three HCV proteins Core, E1, and E2. VSV-HCV-C/E1/E2 is able to replicate efficiently and express the HCV proteins to high levels. In addition, the envelope glycoproteins, E1 and E2, form non-covalently bonded heterodimers that associate in a conformationally authentic manner. They are recognized by conformation-sensitive, monoclonal antibodies that were isolated from HCV infected individuals, indicating that they may be folded in a similar manner as native hepatitis C virions. Given the structural authenticity of these antigens, we tested VSV-HCV-C/E1/E2's ability to stimulate an immune response to the HCV proteins in mice. Our results indicate that both humoral and cellular anti-HCV immune responses were generated to the structural proteins.In an attempt to attenuate the live HCV recombinant VSV vaccine, we genetically deleted the G glycoprotein from the VSV genome (VSVDeltaG-C/E1/E2). This new replication defective virus is unable to produce infectious virus but can still express the HCV structural genes at comparable levels to replication competent virus. When used to immunize mice, VSVDeltaG-C/E1/E2 was able to elicit both Core and E2 specific antibodies, as well as HCV-specific CD 8+ T cells. Collectively, our data indicate that VSV expressing HCV proteins may be a valuable vaccine strategy for hepatitis C infection.

Keywords

Biology, Microbiology; Health Sciences, Immunology

Link to Full Text

http://access.library.miami.edu/login?url=http://gateway.proquest.com/openurl?url_ver=Z39.88-2004&rft_val_fmt=info:ofi/fmt:kev:mtx:dissertation&res_dat=xri:pqdiss&rft_dat=xri:pqdiss:3168710