Cellular immunogenicity of a multi-epitope peptide vaccine candidate based on hepatitis C virus NS5A, NS4B and core proteins in HHD-2 mice

CIGB-230, a mixture of a DNA plasmid expressing hepatitis C virus (HCV) structural antigens and a HCV recombinant capsid protein, has demonstrated to elicit strong immune responses in animals. The present study evaluated the plasmid biodistribution after the administration of CIGB-230 in mice, as well as toxicity of this vaccine candidate in rats. In the biodistribution study, mice received single or repeated intramuscular injections of CIGB-230, 50 μg of plasmid DNA mixed with 5 μg of Co.120 protein. Plasmid presence was assessed in ovaries, kidney, liver, pancreas, mesenteric ganglion, blood, and muscle of the injection site by a qualitative polymerase chain reaction. The toxicology evaluation included treatment groups receiving doses 5, 15, or 50 times higher, according to the body weight, than the expected therapeutic clinical dose. During the first hour after repeated inoculation, a promiscuous distribution was observed. However, 3 months later, plasmid could not be detected in any tissue. There was an absence of detectable adverse effects on key toxicology parameters and no damage evidenced in inspected organs and tissues. These results indicate that CIGB-230 is nontoxic at local and systemic levels and no concerns about persistence are observed, which support clinical testing of this vaccine candidate against HCV.

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High, Broad, Polyfunctional, and Durable T Cell Immune Responses Induced in Mice by a Novel Hepatitis C Virus (HCV) Vaccine Candidate (MVA-HCV) Based on Modified Vaccinia Virus Ankara Expressing the Nearly Full-Length HCV Genome

Journal of Virology ◽

10.1128/jvi.03246-12 ◽

2013 ◽

Vol 87 (13) ◽

pp. 7282-7300 ◽

Cited By ~ 24

Author(s):

C. E. Gomez ◽

B. Perdiguero ◽

M. V. Cepeda ◽

L. Mingorance ◽

J. Garcia-Arriaza ◽

...

Keyword(s):

Hepatitis C Virus ◽

Hepatitis C ◽

T Cell ◽

Immune Responses ◽

Vaccinia Virus ◽

Vaccine Candidate ◽

Full Length ◽

Modified Vaccinia Virus Ankara ◽

T Cell Immune Responses

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Proteasome Activator PA28γ-Dependent Nuclear Retention and Degradation of Hepatitis C Virus Core Protein

Journal of Virology ◽

10.1128/jvi.77.19.10237-10249.2003 ◽

2003 ◽

Vol 77 (19) ◽

pp. 10237-10249 ◽

Cited By ~ 116

Author(s):

Kohji Moriishi ◽

Tamaki Okabayashi ◽

Kousuke Nakai ◽

Kyoji Moriya ◽

Kazuhiko Koike ◽

...

Keyword(s):

Hepatitis C Virus ◽

Hepatitis C ◽

Core Protein ◽

Regulatory Protein ◽

Hcv Infection ◽

Nuclear Retention ◽

Proteasome Activator ◽

Hcv Core Protein ◽

Core Proteins ◽

Chronic Hcv

ABSTRACT Hepatitis C virus (HCV) core protein plays an important role in the formation of the viral nucleocapsid and a regulatory protein involved in hepatocarcinogenesis. In this study, we have identified proteasome activator PA28γ (11S regulator γ) as an HCV core binding protein by using yeast two-hybrid system. This interaction was demonstrated not only in cell culture but also in the livers of HCV core transgenic mice. These findings are extended to human HCV infection by the observation of this interaction in liver specimens from a patient with chronic HCV infection. Neither the interaction of HCV core protein with other PA28 subtypes nor that of PA28γ with other Flavivirus core proteins was detected. Deletion of the PA28γ-binding region from the HCV core protein or knockout of the PA28γ gene led to the export of the HCV core protein from the nucleus to the cytoplasm. Overexpression of PA28γ enhanced the proteolysis of the HCV core protein. Thus, the nuclear retention and stability of the HCV core protein is regulated via a PA28γ-dependent pathway through which HCV pathogenesis may be exerted.

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Sequence heterogeneity of NS5A and core proteins of hepatitis C virus and virological responses to pegylated-interferon/ribavirin combination therapy

Microbiology and Immunology ◽

10.1111/j.1348-0421.2011.00331.x ◽

2011 ◽

Vol 55 (6) ◽

pp. 418-426 ◽

Cited By ~ 13

Author(s):

Ahmed El-Shamy ◽

Ikuo Shoji ◽

Takafumi Saito ◽

Hisayoshi Watanabe ◽

Yoshi-Hiro Ide ◽

...

Keyword(s):

Hepatitis C Virus ◽

Hepatitis C ◽

Combination Therapy ◽

Pegylated Interferon ◽

Sequence Heterogeneity ◽

Core Proteins

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Visualization of Double-Stranded RNA in Cells Supporting Hepatitis C Virus RNA Replication

Journal of Virology ◽

10.1128/jvi.01565-07 ◽

2007 ◽

Vol 82 (5) ◽

pp. 2182-2195 ◽

Cited By ~ 126

Author(s):

Paul Targett-Adams ◽

Steeve Boulant ◽

John McLauchlan

Keyword(s):

Hepatitis C Virus ◽

Hepatitis C ◽

Rna Virus ◽

Three Dimensional ◽

Rna Replication ◽

Hcv Rna ◽

Double Stranded Rna ◽

Core Proteins ◽

Virus Rna ◽

Infected Cells

ABSTRACT The mechanisms involved in hepatitis C virus (HCV) RNA replication are unknown, and this aspect of the virus life cycle is not understood. It is thought that virus-encoded nonstructural proteins and RNA genomes interact on rearranged endoplasmic reticulum (ER) membranes to form replication complexes, which are believed to be sites of RNA synthesis. We report that, through the use of an antibody specific for double-stranded RNA (dsRNA), dsRNA is readily detectable in Huh-7 cells that contain replicating HCV JFH-1 genomes but is absent in control cells. Therefore, as that of other RNA virus genomes, the replication of the HCV genome may involve the generation of a dsRNA replicative intermediate. In Huh-7 cells supporting HCV RNA replication, dsRNA was observed as discrete foci, associated with virus-encoded NS5A and core proteins and identical in morphology and distribution to structures containing HCV RNA visualized by fluorescence-based hybridization methods. Three-dimensional reconstruction of deconvolved z-stack images of virus-infected cells provided detailed insight into the relationship among dsRNA foci, NS5A, the ER, and lipid droplets (LDs). This analysis revealed that dsRNA foci were located on the surface of the ER and often surrounded, partially or wholly, by a network of ER-bound NS5A protein. Additionally, virus-induced dsRNA foci were juxtaposed to LDs, attached to the ER. Thus, we report the visualization of HCV-induced dsRNA foci, the likely sites of virus RNA replication, and propose that HCV genome synthesis occurs at LD-associated sites attached to the ER in virus-infected cells.

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