scholarly journals Expression of HCV Structural Proteins Impairs IFN-Mediated Antiviral Response

Virology ◽  
2002 ◽  
Vol 299 (2) ◽  
pp. 164-171 ◽  
Author(s):  
Päivi Keskinen ◽  
Krister Melén ◽  
Ilkka Julkunen
Keyword(s):  
Antiviral Response ◽  
Structural Proteins ◽  
Hcv Structural Proteins

HCV structural proteins interfere with interferon-alpha Jak/STAT signalling pathway

2007 ◽  
Vol 76 (2) ◽  
pp. 194-197 ◽  
Author(s):  
Esther Luquin ◽  
Esther Larrea ◽  
Maria P. Civeira ◽  
Jesús Prieto ◽  
Rafael Aldabe
Keyword(s):  
Interferon Alpha ◽  
Signalling Pathway ◽  
Structural Proteins ◽  
Hcv Structural Proteins

scholarly journals Evaluating Replication-Defective Vesicular Stomatitis Virus as a Vaccine Vehicle

2006 ◽  
Vol 80 (14) ◽  
pp. 6993-7008 ◽  
Author(s):  
Ayaz M. Majid ◽  
Heather Ezelle ◽  
Sangeeta Shah ◽  
Glen N. Barber
Keyword(s):  
Vesicular Stomatitis Virus ◽  
Cell Activity ◽  
Recombinant Vaccinia Virus ◽  
Antibody Responses ◽  
Structural Proteins ◽  
Effective Vaccine ◽  
Murine Models ◽  
Vesicular Stomatitis ◽  
Ifn Γ ◽  
Hcv Structural Proteins

ABSTRACT We have generated replication-competent (VSV-C/E1/E2) and nonpropagating (VSVΔG-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 VSVΔG-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 VSVΔG-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-γ)-producing CD8+ T cells, as determined by enzyme-linked immunospot (ELISPOT) analysis. Mice immunized with VSVΔG-C/E1/E2 were also protected against the formation of tumors expressing HCV E2 (CT26-hghE2t) and exhibited CT26-hghE2t-specific IFN-γ-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 VSVΔG-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.

scholarly journals Genetic Immunization of Wild-Type and Hepatitis C Virus Transgenic Mice Reveals a Hierarchy of Cellular Immune Response and Tolerance Induction against Hepatitis C Virus Structural Proteins

2001 ◽  
Vol 75 (24) ◽  
pp. 12121-12127 ◽  
Author(s):  
Jujin Satoi ◽  
Kazumoto Murata ◽  
Martin Lechmann ◽  
Elanchezhiyan Manickan ◽  
Zhensheng Zhang ◽  
...  
Keyword(s):  
Immune Response ◽  
Hepatitis C Virus ◽  
Hepatitis C ◽  
Transgenic Mice ◽  
Structural Proteins ◽  
Dna Immunization ◽  
Wild Type ◽  
Genetic Immunization ◽  
Cellular Immune ◽  
Hcv Structural Proteins

ABSTRACT To study the effect of genetic immunization on transgenic expression of hepatitis C virus (HCV) proteins, we evaluated the immunological response of HCV transgenic mice to HCV expression plasmids. FVB/n transgenic mice expressing HCV structural proteins (core, E1, and E2) and wild-type (WT) FVB/n mice were immunized intramuscularly with plasmids expressing core (pHCVcore) or core/E1/E2 (pHCVSt). After immunization, HCV-specific humoral and cellular immune response was studied. Both WT and transgenic mice immunized with either HCV construct produced antibodies and exhibited T-cell proliferative responses against core or envelope. In WT mice immunized with pHCVSt, cytotoxic T-lymphocyte (CTL) activities were detected against E2 but not against core or E1, whereas strong CTL activities against core could be detected in WT mice immunized with pHCVcore. In pHCVSt-immunized, transgenic mice, CTL activities against the core or envelope were completely absent, but core-specific CTL activities could be detected in pHCVcore-immunized transgenic mice. A similar pattern of immune responses was also observed in other mouse strains, including a transgenic line expressing human HLA-A2.1 molecules (AAD mice). Despite the presence of a peripheral cellular immunity against HCV, no liver pathology or lymphocytic infiltrate was observed in these transgenic mice. Our study suggests a hierarchy of CTL response against the HCV structural proteins (E2 > core > E1) in vivo when the proteins are expressed as a polyprotein. The HCV transgenic mice can be induced by DNA immunization to generate anti-HCV antibodies and anticore CTLs. However, they are tolerant at the CTL level against the E2 protein despite DNA immunization.

scholarly journals Hepatitis C Virus Structural Proteins Assemble into Viruslike Particles in Insect Cells

1998 ◽  
Vol 72 (5) ◽  
pp. 3827-3836 ◽  
Author(s):  
Thomas F. Baumert ◽  
Susumu Ito ◽  
David T. Wong ◽  
T. Jake Liang
Keyword(s):  
Hepatitis C Virus ◽  
Hepatitis C ◽  
Insect Cells ◽  
Vaccine Development ◽  
Cultured Cells ◽  
Recombinant Baculovirus ◽  
Structural Proteins ◽  
Particle Assembly ◽  
Viruslike Particles ◽  
Hcv Structural Proteins

ABSTRACT Hepatitis C virus (HCV) is a leading cause of chronic hepatitis in the world. The study of HCV has been hampered by the low level of viral particles in infected individuals, the inability to propagate efficiently the virus in cultured cells, and the lack of a convenient animal model. Due to these obstacles, neither the structure of the virus nor the prerequisites for its assembly have been clearly defined. In this report, we describe a model for the production and purification of HCV-like particles in insect cells using a recombinant baculovirus containing the cDNA of the HCV structural proteins. In insect cells, expressed HCV structural proteins assembled into enveloped viruslike particles (40 to 60 nm in diameter) in large cytoplasmic cisternae, presumably derived from the endoplasmic reticulum. Biophysical characterization of viruslike particles by CsCl and sucrose gradient centrifugation revealed biophysical properties similar to those of putative virions isolated from infected humans. The results suggested that HCV core and envelope proteins without p7 were sufficient for viral particle formation. Analysis of particle-associated nucleic acids demonstrated that HCV RNAs were selectively incorporated into the particles over non-HCV transcripts. The synthesis of HCV-like particles in insect cells may provide an important tool to determine the structural requirements for HCV particle assembly as well as to study viral genome encapsidation and virus-host interactions. The described system may also represent a potential approach toward vaccine development.

scholarly journals Generation of Hepatitis C Virus-Like Particles by Use of a Recombinant Vesicular Stomatitis Virus Vector

2002 ◽  
Vol 76 (23) ◽  
pp. 12325-12334 ◽  
Author(s):  
Heather J. Ezelle ◽  
Dubravka Markovic ◽  
Glen N. Barber
Keyword(s):  
Hepatitis C Virus ◽  
Hepatitis C ◽  
Vesicular Stomatitis Virus ◽  
Etiologic Agent ◽  
Hcv Infection ◽  
Structural Proteins ◽  
Coding Region ◽  
Vesicular Stomatitis ◽  
Hcv Structural Proteins

ABSTRACT Hepatitis C virus (HCV), a major etiologic agent of hepatocellular carcinoma, presently infects approximately 400 million people worldwide, making the development of protective measures against HCV infection a key objective. Here we have generated a recombinant vesicular stomatitis virus (VSV), which expresses the HCV structural proteins, by inserting the contiguous Core, E1, and E2 coding region of HCV into the VSV genome. Recombinant VSV expressing HCV Core, E1, and E2 (VSV-HCV-C/E1/E2) grew to high titers in vitro and efficiently expressed the incorporated HCV gene product, which became fully processed into the individual HCV structural proteins. Biochemical and biophysical analysis indicated that the HCV Core, E1, and E2 proteins assembled to form HCV-like particles (HCV-LPs) possessing properties similar to the ultrastructural properties of HCV virions. Mice immunized with VSV-HCV-C/E1/E2 generated cell-mediated immune responses to all of the HCV structural proteins, and humoral responses, particularly to E2, were also readily evident. Our data collectively indicate that engineered VSVs expressing HCV Core, E1, and E2 and/or HCV-LPs represent useful tools in vaccine and immunotherapeutic strategies designed to address HCV infection.

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