scholarly journals Assembly of Hepatitis C Virus Antigens Expressed from a Recombinant Vaccinia Virus.

1996 ◽  
Vol 72 (4) ◽  
pp. 73-78 ◽  
Author(s):  
Shigeru YOSHIDA ◽  
Sadao MANABE ◽  
Osamu TANISHITA ◽  
Chie KAJI ◽  
Yasuyuki GOMI ◽  
...  
Keyword(s):  
Hepatitis C Virus ◽  
Hepatitis C ◽  
Vaccinia Virus ◽  
Recombinant Vaccinia Virus ◽  
Recombinant Vaccinia ◽  
Virus Antigens

scholarly journals Development of Cell-Based Assays for In Vitro Characterization of Hepatitis C Virus NS3/4A Protease Inhibitors

2005 ◽  
Vol 49 (4) ◽  
pp. 1381-1390 ◽  
Author(s):  
Victoria Chung ◽  
Anthony R. Carroll ◽  
Norman M. Gray ◽  
Nigel R. Parry ◽  
Pia A. Thommes ◽  
...  
Keyword(s):  
Hepatitis C Virus ◽  
Hepatitis C ◽  
Vaccinia Virus ◽  
Protease Inhibitors ◽  
Expression System ◽  
Recombinant Vaccinia Virus ◽  
Enzyme Linked Immunosorbent Assay ◽  
Recombinant Vaccinia

ABSTRACT A recombinant vaccinia virus, expressing the NS3-to-NS5 region of the N clone of hepatitis C virus (HCV), was generated and utilized both in a gel-based assay and in an enzyme-linked immunosorbent assay (ELISA) to evaluate the pyrrolidine-5,5-trans-lactams, a series of inhibitors of the HCV NS3/4A protease. The absolute levels of processed, mature HCV nonstructural proteins in this system were found to decrease in the presence of the trans-lactams. Monitoring of this reduction enabled end points and 50% inhibitory concentrations to be calculated in order to rank the active compounds according to potency. These compounds had no effect on the transcription or translation of the NS3-5 polyprotein at concentrations shown to inhibit NS3/4A protease, and they were shown to be specific inhibitors of this protease. The ELISA, originally developed using the vaccinia virus expression system, was modified to utilize Huh-7 cells containing an HCV replicon. Results with this assay correlated well with those obtained with the recombinant vaccinia virus assays. These results demonstrate the utility of these assays for the characterization of NS3/4A protease inhibitors. In addition, inhibitors of other viral targets, such as polymerase and helicase, can be evaluated in the context of the replicon ELISA.

Vaccination with an adenoviral vector encoding hepatitis C virus (HCV) NS3 protein protects against infection with HCV-recombinant vaccinia virus

Vaccine ◽  
2002 ◽  
Vol 21 (3-4) ◽  
pp. 202-210 ◽  
Author(s):  
Laura Arribillaga ◽  
Ascensión López Dı́az de Cerio ◽  
Pablo Sarobe ◽  
Noelia Casares ◽  
Marta Gorraiz ◽  
...  
Keyword(s):  
Hepatitis C Virus ◽  
Hepatitis C ◽  
Vaccinia Virus ◽  
Adenoviral Vector ◽  
Recombinant Vaccinia Virus ◽  
Recombinant Vaccinia ◽  
Ns3 Protein

scholarly journals Expression and characterization of glycoprotein gp35 of hepatitis C virus using recombinant vaccinia virus

1992 ◽  
Vol 73 (9) ◽  
pp. 2313-2318 ◽  
Author(s):  
M. Kohara ◽  
K. Tsukiyama-Kohara ◽  
N. Maki ◽  
K. Asano ◽  
K. Yamaguchi ◽  
...  
Keyword(s):  
Hepatitis C Virus ◽  
Hepatitis C ◽  
Vaccinia Virus ◽  
Recombinant Vaccinia Virus ◽  
Recombinant Vaccinia

scholarly journals Dendritic cells pulsed with hepatitis C virus NS3 protein induce immune responses and protection from infection with recombinant vaccinia virus expressing NS3

2006 ◽  
Vol 87 (1) ◽  
pp. 1-10 ◽  
Author(s):  
Hong Yu ◽  
Hui Huang ◽  
Jim Xiang ◽  
Lorne A. Babiuk ◽  
Sylvia van Drunen Littel-van den Hurk
Keyword(s):  
Dendritic Cells ◽  
Hepatitis C Virus ◽  
Hepatitis C ◽  
Immune Responses ◽  
Vaccinia Virus ◽  
Recombinant Vaccinia Virus ◽  
Interleukin 12 ◽  
Cpg Odn ◽  
Recombinant Vaccinia ◽  
Ns3 Protein

Infections with Hepatitis C virus (HCV) pose a serious health problem worldwide. In this study, the hypothesis that adoptive transfer of dendritic cells (DCs) pulsed with HCV NS3 protein and matured with an oligodeoxynucleotide (ODN) containing CpG motifs (CpG) ex vivo would initiate potent HCV-specific protective immune responses in vivo was tested. NS3 protein was efficiently transduced into DCs and treatment of DCs with CpG ODN induced phenotypic maturation and specifically increased the expression of CD40. DCs matured with CpG ODN produced higher interleukin 12 levels and a stronger allogeneic T-cell response compared with untreated DCs. Notably, there were no differences between NS3-pulsed DCs and DCs pulsed with a control protein with respect to phenotype, cytokine production or mixed lymphocyte reaction, indicating that transduction with NS3 protein did not impair DC functions. Compared with the untreated NS3-pulsed DCs, the NS3-pulsed DCs matured with CpG ODN induced stronger cellular immune responses including enhanced cytotoxicity, higher interferon-γ production and stronger lymphocyte proliferation. Upon challenge with a recombinant vaccinia virus expressing NS3, all mice immunized with NS3-pulsed DCs showed a significant reduction in vaccinia virus titres when compared with mock-immunized mice. However, the NS3-pulsed DCs matured with CpG ODN induced higher levels of protection compared with the untreated NS3-pulsed DCs. These data are the first to show that NS3-pulsed DCs induce specific immune responses and provide protection from viral challenge, and also demonstrate that CpG ODNs, which have a proven safety profile, would be useful in the development of DC vaccines.

scholarly journals Antisense Oligonucleotide Inhibition of Hepatitis C Virus (HCV) Gene Expression in Livers of Mice Infected with an HCV-Vaccinia Virus Recombinant

1999 ◽  
Vol 43 (2) ◽  
pp. 347-353 ◽  
Author(s):  
Hong Zhang ◽  
Ronnie Hanecak ◽  
Vickie Brown-Driver ◽  
Raana Azad ◽  
Boyd Conklin ◽  
...  
Keyword(s):  
Gene Expression ◽  
Animal Model ◽  
Hepatitis C Virus ◽  
Hepatitis C ◽  
Vaccinia Virus ◽  
Antisense Oligonucleotides ◽  
Small Animal ◽  
Initiation Codon ◽  
Small Animal Model ◽  
Recombinant Vaccinia

ABSTRACT Hepatitis C virus (HCV) is the major cause of non-A, non-B hepatitis worldwide. Current treatments are not curative for most infected individuals, and there is an urgent need for both novel therapeutic agents and small-animal models which can be used to evaluate candidate drugs. A small-animal model of HCV gene expression was developed with recombinant vaccinia virus vectors. VHCV-IRES (internal ribosome entry site) is a recombinant vaccinia viral vector containing the HCV 5′ nontranslated region (5′-NTR) and a portion of the HCV core coding region fused to the firefly luciferase gene. Intraperitoneal injection of VHCV-IRES produced high levels of luciferase activity in the livers of BALB/c mice. Antisense oligonucleotides complementary to the HCV 5′-NTR and translation initiation codon regions were then evaluated for their effects on the expression of these target HCV sequences in BALB/c mice infected with the vaccinia virus vector. Treatment of VHCV-IRES-infected mice with 20-base phosphorothioate oligonucleotides complementary to the sequence surrounding the HCV initiation codon (nucleotides 330 to 349) specifically reduced luciferase expression in the livers in a dose-dependent manner. Inhibition of HCV reporter gene expression in this small-animal model suggests that antisense oligonucleotides may provide a novel therapy for treatment of chronic HCV infection.

Antibody responses and protective immunity to recombinant vaccinia virus-expressed bluetongue virus antigens

1997 ◽  
Vol 59 (3-4) ◽  
pp. 293-309 ◽  
Author(s):  
Zelia I.P. Lobato ◽  
Barbara E.H. Coupar ◽  
Christian P. Gray ◽  
Ross Lunt ◽  
Marion E. Andrew
Keyword(s):  
Vaccinia Virus ◽  
Bluetongue Virus ◽  
Protective Immunity ◽  
Recombinant Vaccinia Virus ◽  
Antibody Responses ◽  
Recombinant Vaccinia ◽  
Virus Antigens
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