start-ver=1.4
cd-journal=joma
no-vol=291
cd-vols=
no-issue=6
article-no=
start-page=1119
end-page=1130
dt-received=
dt-revised=
dt-accepted=
dt-pub-year=2023
dt-pub=20231020
dt-online=
en-article=
kn-article=
en-subject=
kn-subject=
en-title=
kn-title=Hepatitis C virus NS5B triggers an MDA5-mediated innate immune response by producing dsRNA without the replication of viral genomes
en-subtitle=
kn-subtitle=
en-abstract=
kn-abstract=During the replication of viral genomes, RNA viruses produce double-stranded RNA (dsRNA), through the activity of their RNA-dependent RNA polymerases (RdRps) as viral replication intermediates. Recognition of viral dsRNA by host pattern recognition receptors – such as retinoic acid-induced gene-I (RIG-I)-like receptors and Toll-like receptor 3 – triggers the production of interferon (IFN)-β via the activation of IFN regulatory factor (IRF)-3. It has been proposed that, during the replication of viral genomes, each of RIG-I and melanoma differentiation-associated gene 5 (MDA5) form homodimers for the efficient activation of a downstream signalling pathway in host cells. We previously reported that, in the non-neoplastic human hepatocyte line PH5CH8, the RdRp NS5B derived from hepatitis C virus (HCV) could induce IFN-β expression by its RdRp activity without the actual replication of viral genomes. However, the exact mechanism by which HCV NS5B produced IFN-β remained unknown. In the present study, we first showed that NS5B derived from another Flaviviridae family member, GB virus B (GBV-B), also possessed the ability to induce IFN-β in PH5CH8 cells. Similarly, HCV NS5B, but not its G317V mutant, which lacks RdRp activity, induced the dimerization of MDA5 and subsequently the activation of IRF-3. Interestingly, immunofluorescence analysis showed that HCV NS5B produced dsRNA. Like HCV NS5B, GBV-B NS5B also triggered the production of dsRNA and subsequently the dimerization of MDA5. Taken together, our results show that HCV NS5B triggers an MDA5-mediated innate immune response by producing dsRNA without the replication of viral genomes in human hepatocytes.
en-copyright=
kn-copyright=
en-aut-name=DansakoHiromichi
en-aut-sei=Dansako
en-aut-mei=Hiromichi
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=1
ORCID=
en-aut-name=IkedaMasanori
en-aut-sei=Ikeda
en-aut-mei=Masanori
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=2
ORCID=
en-aut-name=AriumiYasuo
en-aut-sei=Ariumi
en-aut-mei=Yasuo
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=3
ORCID=
en-aut-name=TogashiYosuke
en-aut-sei=Togashi
en-aut-mei=Yosuke
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=4
ORCID=
en-aut-name=KatoNobuyuki
en-aut-sei=Kato
en-aut-mei=Nobuyuki
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=5
ORCID=
affil-num=1
en-affil=Department of Tumor Microenvironment, Faculty of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University
kn-affil=
affil-num=2
en-affil=Division of Biological Information Technology, Joint Research Center for Human Retrovirus Infection, Kagoshima University
kn-affil=
affil-num=3
en-affil=Management Department of Biosafety, Laboratory Animal, and Pathogen Bank, National Institute of Infectious Diseases
kn-affil=
affil-num=4
en-affil=Department of Tumor Microenvironment, Faculty of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University
kn-affil=
affil-num=5
en-affil=Department of Tumor Microenvironment, Faculty of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University
kn-affil=
en-keyword=double-stranded RNA
kn-keyword=double-stranded RNA
en-keyword=hepatitis C virus
kn-keyword=hepatitis C virus
en-keyword=innate immunity
kn-keyword=innate immunity
en-keyword=RIG-I-like receptor
kn-keyword=RIG-I-like receptor
en-keyword=RNA virus
kn-keyword=RNA virus
END
start-ver=1.4
cd-journal=joma
no-vol=132
cd-vols=
no-issue=3
article-no=
start-page=131
end-page=143
dt-received=
dt-revised=
dt-accepted=
dt-pub-year=2020
dt-pub=20201201
dt-online=
en-article=
kn-article=
en-subject=
kn-subject=
en-title=Hepatitis C virus (HCV) : Development of anti-HCV agents and anti-HCV therapy
kn-title=Ｃ型肝炎ウイルス（HCV）：抗 HCV 剤の開発と抗 HCV 療法
en-subtitle=
kn-subtitle=
en-abstract=
kn-abstract=
en-copyright=
kn-copyright=
en-aut-name=KatoNobuyuki
en-aut-sei=Kato
en-aut-mei=Nobuyuki
kn-aut-name=加藤宣之
kn-aut-sei=加藤
kn-aut-mei=宣之
aut-affil-num=1
ORCID=
affil-num=1
en-affil=Department of Tumor Virology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences
kn-affil=岡山大学大学院医歯薬学総合研究科 腫瘍ウイルス学
en-keyword=インターフェロン
kn-keyword=インターフェロン
en-keyword=リバビリン
kn-keyword=リバビリン
en-keyword=HCVレプリコンシステム
kn-keyword=HCVレプリコンシステム
en-keyword=抗HCVアッセイシステム
kn-keyword=抗HCVアッセイシステム
en-keyword= DAA
kn-keyword= DAA
END
start-ver=1.4
cd-journal=joma
no-vol=132
cd-vols=
no-issue=2
article-no=
start-page=60
end-page=67
dt-received=
dt-revised=
dt-accepted=
dt-pub-year=2020
dt-pub=20200803
dt-online=
en-article=
kn-article=
en-subject=
kn-subject=
en-title=Hepatitis C virus (HCV)：Diversity and variation of RNA genome
kn-title=C型肝炎ウイルス（HCV）：RNAゲノムの多様性と変異性
en-subtitle=
kn-subtitle=
en-abstract=
kn-abstract=
en-copyright=
kn-copyright=
en-aut-name=KatoNobuyuki
en-aut-sei=Kato
en-aut-mei=Nobuyuki
kn-aut-name=加藤宣之
kn-aut-sei=加藤
kn-aut-mei=宣之
aut-affil-num=1
ORCID=
affil-num=1
en-affil=Department of Tumor Virology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences
kn-affil=岡山大学大学院医歯薬学総合研究科 腫瘍ウイルス学
en-keyword=HCV ゲノム
kn-keyword=HCV ゲノム
en-keyword=レプリコン複製細胞
kn-keyword=レプリコン複製細胞
en-keyword=長期継代培養
kn-keyword=長期継代培養
en-keyword=遺伝子解析
kn-keyword=遺伝子解析
en-keyword=準種
kn-keyword=準種
END
start-ver=1.4
cd-journal=joma
no-vol=93
cd-vols=
no-issue=7
article-no=
start-page=1422
end-page=1431
dt-received=
dt-revised=
dt-accepted=
dt-pub-year=2012
dt-pub=201207
dt-online=
en-article=
kn-article=
en-subject=
kn-subject=
en-title=
kn-title=Development of hepatitis C virus production reporter-assay systems using two different hepatoma cell lines
en-subtitle=
kn-subtitle=
en-abstract=
kn-abstract= A hepatitis C virus (HCV) infection system was developed previously using the HCV JFH-1 strain (genotype 2a) and HuH-7 cells, and this cell culture is so far the only robust production system for HCV. In patients with chronic hepatitis C, the virological effects of pegylated interferon and ribavirin therapy differ depending on the HCV strain and the genetic background of the host. Recently, we reported the hepatoma-derived Li23 cell line, in which the JFH-1 life cycle is reproduced at a level almost equal to that in HuH-7-derived RSc cells. To monitor the HCV life cycle more easily, we here developed JFH-1 reporter-assay systems using both HuH-7- and Li23-derived cell lines. To identify any genetic mutations by long-term cell culture, HCV RNAs in HuH-7 cells were amplified 130 days after infection and subjected to sequence analysis to find adaptive mutation(s) for robust virus replication. We identified two mutations, H2505Q and V2995L, in the NS5B region. V2995L but not H2505Q enhanced JFH-1 RNA replication. However, we found that H2505Q but not V2995L enhanced HCV RNA replication of strain O (genotype 1b). We also selected highly permissive D7 cells by serial subcloning of Li23 cells. The expression levels of claudin-1 and Niemann-Pick C1-like 1 in D7 cells are higher than those in parental Li23 cells. In this study, we developed HCV JFH-1 reporter-assay systems using two distinct hepatoma cell lines, HuH-7 and Li23. The mutations in NS5B resulted in different effects on strains O and JFH-1 HCV RNA replication.
en-copyright=
kn-copyright=
en-aut-name=TakedaMidori
en-aut-sei=Takeda
en-aut-mei=Midori
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=1
ORCID=
en-aut-name=IkedaMasanori
en-aut-sei=Ikeda
en-aut-mei=Masanori
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=2
ORCID=
en-aut-name=AriumiYasuo
en-aut-sei=Ariumi
en-aut-mei=Yasuo
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=3
ORCID=
en-aut-name=WakitaTakaji
en-aut-sei=Wakita
en-aut-mei=Takaji
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=4
ORCID=
en-aut-name=KatoNobuyuki
en-aut-sei=Kato
en-aut-mei=Nobuyuki
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=5
ORCID=
affil-num=1
en-affil=Department of Tumor Virology, Okayama University, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences
kn-affil=
affil-num=2
en-affil=Department of Tumor Virology, Okayama University, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences
kn-affil=
affil-num=3
en-affil=Department of Tumor Virology, Okayama University, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences
kn-affil=
affil-num=4
en-affil=Department of Virology II, National Institute of Infectious Disease
kn-affil=
affil-num=5
en-affil=Department of Tumor Virology, Okayama University, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences
kn-affil=
END
start-ver=1.4
cd-journal=joma
no-vol=167
cd-vols=
no-issue=1
article-no=
start-page=74
end-page=85
dt-received=
dt-revised=
dt-accepted=
dt-pub-year=2012
dt-pub=201207
dt-online=
en-article=
kn-article=
en-subject=
kn-subject=
en-title=
kn-title=Identification of host genes showing differential expression profiles with cell-based long-term replication of hepatitis C virus RNA
en-subtitle=
kn-subtitle=
en-abstract=
kn-abstract= Persistent hepatitis C virus (HCV) infection frequently causes hepatocellular carcinoma. However, the mechanisms of HCV-associated hepatocarcinogenesis and disease progression are unclear. Although the human hepatoma cell line, HuH-7, has been widely used as the only cell culture system for robust HCV replication, we recently developed new human hepatoma Li23 cell line-derived OL, OL8, OL11, and OL14 cells, in which genome-length HCV RNA (O strain of genotype 1b) efficiently replicates. OL, OL8, OL11, and OL14 cells were cultured for more than 2 years. We prepared cured cells from OL8 and OL11 cells by interferon-γ treatment. The cured cells were also cultured for more than 2 years. cDNA microarray and RT-PCR analyses were performed using total RNAs prepared from these cells. We first selected several hundred highly or moderately expressed probes, the expression levels of which were upregulated or downregulated at ratios of more than 2 or less than 0.5 in each set of compared cells (e.g., parent OL8 cells versus OL8 cells cultured for 2 years). From among these probes, we next selected those whose expression levels commonly changed during a 2-year culture of genome-length HCV RNA-replicating cells, but which did not change during a 2-year culture period in cured cells. We further examined the expression levels of the selected candidate genes by RT-PCR analysis using additional specimens from the cells cultured for 3.5 years. Reproducibility of the RT-PCR analysis using specimens from recultured cells was also confirmed. Finally, we identified 5 upregulated genes and 4 downregulated genes, the expression levels of which were irreversibly altered during 3.5-year replication of HCV RNA. These genes may play roles in the optimization of the environment in HCV RNA replication, or may play key roles in the progression of HCV-associated hepatic diseases.
en-copyright=
kn-copyright=
en-aut-name=SejimaHiroe
en-aut-sei=Sejima
en-aut-mei=Hiroe
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=1
ORCID=
en-aut-name=MoriKyoko
en-aut-sei=Mori
en-aut-mei=Kyoko
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=2
ORCID=
en-aut-name=AriumiYasuo
en-aut-sei=Ariumi
en-aut-mei=Yasuo
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=3
ORCID=
en-aut-name=IkedaMasanori
en-aut-sei=Ikeda
en-aut-mei=Masanori
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=4
ORCID=
en-aut-name=KatoNobuyuki
en-aut-sei=Kato
en-aut-mei=Nobuyuki
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=5
ORCID=
affil-num=1
en-affil=Department of Tumor Virology, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences
kn-affil=
affil-num=2
en-affil=Department of Tumor Virology, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences
kn-affil=
affil-num=3
en-affil=Department of Tumor Virology, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences
kn-affil=
affil-num=4
en-affil=Department of Tumor Virology, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences
kn-affil=
affil-num=5
en-affil=Department of Tumor Virology, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences
kn-affil=
en-keyword=HCV
kn-keyword=HCV
en-keyword=HCV RNA replication system
kn-keyword=HCV RNA replication system
en-keyword=Li23 cells
kn-keyword=Li23 cells
en-keyword=Long-term RNA replication
kn-keyword=Long-term RNA replication
en-keyword=Upregulated host genes
kn-keyword=Upregulated host genes
en-keyword=Downregulated host genes
kn-keyword=Downregulated host genes
END
start-ver=1.4
cd-journal=joma
no-vol=9
cd-vols=
no-issue=3
article-no=
start-page=e91156
end-page=
dt-received=
dt-revised=
dt-accepted=
dt-pub-year=2014
dt-pub=20140313
dt-online=
en-article=
kn-article=
en-subject=
kn-subject=
en-title=
kn-title=Genetic Characterization of Hepatitis C Virus in Long-Term RNA Replication Using Li23 Cell Culture Systems
en-subtitle=
kn-subtitle=
en-abstract=
kn-abstract=Background
The most distinguishing genetic feature of hepatitis C virus (HCV) is its remarkable diversity and variation. To understand this feature, we previously performed genetic analysis of HCV in the long-term culture of human hepatoma HuH-7-derived HCV RNA-replicating cell lines. On the other hand, we newly established HCV RNA-replicating cell lines using human hepatoma Li23 cells, which were distinct from HuH-7 cells.
Methodology/Principal Findings
Li23-derived HCV RNA-replicating cells were cultured for 4 years. We performed genetic analysis of HCVs recovered from these cells at 0, 2, and 4 years in culture. Most analysis was performed in two separate parts: one part covered from the 5′-terminus to NS2, which is mostly nonessential for RNA replication, and the other part covered from NS3 to NS5B, which is essential for RNA replication. Genetic mutations in both regions accumulated in a time-dependent manner, and the mutation rates in the 5′-terminus-NS2 and NS3-NS5B regions were 4.0–9.0×10−3 and 2.7–4.0×10−3 base substitutions/site/year, respectively. These results suggest that the variation in the NS3-NS5B regions is affected by the pressure of RNA replication. Several in-frame deletions (3–105 nucleotides) were detected in the structural regions of HCV RNAs obtained from 2-year or 4-year cultured cells. Phylogenetic tree analyses clearly showed that the genetic diversity of HCV was expanded in a time-dependent manner. The GC content of HCV RNA was significantly increased in a time-dependent manner, as previously observed in HuH-7-derived cell systems. This phenomenon was partially due to the alterations in codon usages for codon optimization in human cells. Furthermore, we demonstrated that these long-term cultured cells were useful as a source for the selection of HCV clones showing resistance to anti-HCV agents.
Conclusions/Significance
Long-term cultured HCV RNA-replicating cells are useful for the analysis of evolutionary dynamics and variations of HCV and for drug-resistance analysis.
en-copyright=
kn-copyright=
en-aut-name=KatoNobuyuki
en-aut-sei=Kato
en-aut-mei=Nobuyuki
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=1
ORCID=
en-aut-name=SejimaHiroe
en-aut-sei=Sejima
en-aut-mei=Hiroe
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=2
ORCID=
en-aut-name=UedaYouki
en-aut-sei=Ueda
en-aut-mei=Youki
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=3
ORCID=
en-aut-name=MoriKyoko
en-aut-sei=Mori
en-aut-mei=Kyoko
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=4
ORCID=
en-aut-name=SatohShinya
en-aut-sei=Satoh
en-aut-mei=Shinya
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=5
ORCID=
en-aut-name=DansakoHiromichi
en-aut-sei=Dansako
en-aut-mei=Hiromichi
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=6
ORCID=
en-aut-name=IkedaMasanori
en-aut-sei=Ikeda
en-aut-mei=Masanori
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=7
ORCID=
affil-num=1
en-affil=
kn-affil=Department of Tumor Virology, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences
affil-num=2
en-affil=
kn-affil=Department of Tumor Virology, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences
affil-num=3
en-affil=
kn-affil=Department of Tumor Virology, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences
affil-num=4
en-affil=
kn-affil=Department of Tumor Virology, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences
affil-num=5
en-affil=
kn-affil=Department of Tumor Virology, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences
affil-num=6
en-affil=
kn-affil=Department of Tumor Virology, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences
affil-num=7
en-affil=
kn-affil=Department of Tumor Virology, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences
END
start-ver=1.4
cd-journal=joma
no-vol=8
cd-vols=
no-issue=8
article-no=
start-page=e72519
end-page=
dt-received=
dt-revised=
dt-accepted=
dt-pub-year=2013
dt-pub=20130830
dt-online=
en-article=
kn-article=
en-subject=
kn-subject=
en-title=
kn-title=New Preclinical Antimalarial Drugs Potently Inhibit Hepatitis C Virus Genotype 1b RNA Replication
en-subtitle=
kn-subtitle=
en-abstract=
kn-abstract=BACKGROUND:
Persistent hepatitis C virus (HCV) infection causes chronic liver diseases and is a global health problem. Although new triple therapy (pegylated-interferon, ribavirin, and telaprevir/boceprevir) has recently been started and is expected to achieve a sustained virologic response of more than 70% in HCV genotype 1 patients, there are several problems to be resolved, including skin rash/ageusia and advanced anemia. Thus a new type of anti-HCV drug is still needed.
METHODOLOGY/PRINCIPAL FINDINGS:
Recently developed HCV drug assay systems using HCV-RNA-replicating cells (e.g., HuH-7-derived OR6 and Li23-derived ORL8) were used to evaluate the anti-HCV activity of drug candidates. During the course of the evaluation of anti-HCV candidates, we unexpectedly found that two preclinical antimalarial drugs (N-89 and its derivative N-251) showed potent anti-HCV activities at tens of nanomolar concentrations irrespective of the cell lines and HCV strains of genotype 1b. We confirmed that replication of authentic HCV-RNA was inhibited by these drugs. Interestingly, however, this anti-HCV activity did not work for JFH-1 strain of genotype 2a. We demonstrated that HCV-RNA-replicating cells were cured by treatment with only N-89. A comparative time course assay using N-89 and interferon-α demonstrated that N-89-treated ORL8 cells had more rapid anti-HCV kinetics than did interferon-α-treated cells. This anti-HCV activity was largely canceled by vitamin E. In combination with interferon-α and/or ribavirin, N-89 or N-251 exhibited a synergistic inhibitory effect.
CONCLUSIONS/SIGNIFICANCE:
We found that the preclinical antimalarial drugs N-89 and N-251 exhibited very fast and potent anti-HCV activities using cell-based HCV-RNA-replication assay systems. N-89 and N-251 may be useful as a new type of anti-HCV reagents when used singly or in combination with interferon and/or ribavirin.
en-copyright=
kn-copyright=
en-aut-name=UedaYouki
en-aut-sei=Ueda
en-aut-mei=Youki
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=1
ORCID=
en-aut-name=TakedaMidori
en-aut-sei=Takeda
en-aut-mei=Midori
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=2
ORCID=
en-aut-name=MoriKyoko
en-aut-sei=Mori
en-aut-mei=Kyoko
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=3
ORCID=
en-aut-name=DansakoHiromichi
en-aut-sei=Dansako
en-aut-mei=Hiromichi
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=4
ORCID=
en-aut-name=WakitaTakaji
en-aut-sei=Wakita
en-aut-mei=Takaji
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=5
ORCID=
en-aut-name=KimHye-Sook
en-aut-sei=Kim
en-aut-mei=Hye-Sook
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=6
ORCID=
en-aut-name=SatoAkira
en-aut-sei=Sato
en-aut-mei=Akira
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=7
ORCID=
en-aut-name=WatayaYusuke
en-aut-sei=Wataya
en-aut-mei=Yusuke
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=8
ORCID=
en-aut-name=IkedaMasanori
en-aut-sei=Ikeda
en-aut-mei=Masanori
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=9
ORCID=
en-aut-name=KatoNobuyuki
en-aut-sei=Kato
en-aut-mei=Nobuyuki
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=10
ORCID=
affil-num=1
en-affil=
kn-affil=Department of Tumor Virology, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences
affil-num=2
en-affil=
kn-affil=Department of Tumor Virology, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences
affil-num=3
en-affil=
kn-affil=Department of Tumor Virology, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences
affil-num=4
en-affil=
kn-affil=Department of Tumor Virology, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences
affil-num=5
en-affil=
kn-affil=Department of Virology II, National Institute of Infectious Disease
affil-num=6
en-affil=
kn-affil=Department of Drug Informatics, Faculty of Pharmaceutical Sciences, Okayama University
affil-num=7
en-affil=
kn-affil=Department of Drug Informatics, Faculty of Pharmaceutical Sciences, Okayama University
affil-num=8
en-affil=
kn-affil=Department of Drug Informatics, Faculty of Pharmaceutical Sciences, Okayama University
affil-num=9
en-affil=
kn-affil=Department of Tumor Virology, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences
affil-num=10
en-affil=
kn-affil=Department of Tumor Virology, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences
END
start-ver=1.4
cd-journal=joma
no-vol=
cd-vols=
no-issue=
article-no=
start-page=
end-page=
dt-received=
dt-revised=
dt-accepted=
dt-pub-year=2015
dt-pub=2015
dt-online=
en-article=
kn-article=
en-subject=
kn-subject=
en-title=
kn-title=The cyclic GMP-AMP synthetase-STING signaling pathway is required for both the innate immune response against HBV and the suppression of HBV assembly
en-subtitle=
kn-subtitle=
en-abstract=
kn-abstract=During viral replication, the innate immune response is induced through the recognition of viral replication intermediates by host factor(s). One of these host factors, cyclic GMP-AMP synthetase (cGAS), was recently reported to be involved in the recognition of viral DNA derived from DNA viruses. However, it is uncertain whether cGAS is involved in the recognition of hepatitis B virus (HBV), which is a hepatotropic DNA virus. In the present study, we demonstrated that HBV genome-derived dsDNA induced the innate immune response through cGAS and its adaptor protein, STING, in human hepatoma Li23 cells expressing high levels of cGAS. In addition, we demonstrated that HBV infection induced ISG56 through the cGAS-STING signaling pathway. This signaling pathway also showed an antiviral response towards HBV through the suppression of viral assembly. From these results, we conclude that the cGAS-STING signaling pathway is required for not only the innate immune response against HBV but also the suppression of HBV assembly. The cGAS-STING signaling pathway may thus be a novel target for anti-HBV strategies.
en-copyright=
kn-copyright=
en-aut-name=DansakoHiromichi
en-aut-sei=Dansako
en-aut-mei=Hiromichi
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=1
ORCID=
en-aut-name=UedaYouki
en-aut-sei=Ueda
en-aut-mei=Youki
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=2
ORCID=
en-aut-name=OkumuraNobuaki
en-aut-sei=Okumura
en-aut-mei=Nobuaki
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=3
ORCID=
en-aut-name=SatohShinya
en-aut-sei=Satoh
en-aut-mei=Shinya
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=4
ORCID=
en-aut-name=SugiyamaMasaya
en-aut-sei=Sugiyama
en-aut-mei=Masaya
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=5
ORCID=
en-aut-name=MizokamiMasashi
en-aut-sei=Mizokami
en-aut-mei=Masashi
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=6
ORCID=
en-aut-name=IkedaMasanori
en-aut-sei=Ikeda
en-aut-mei=Masanori
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=7
ORCID=
en-aut-name=KatoNobuyuki
en-aut-sei=Kato
en-aut-mei=Nobuyuki
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=8
ORCID=
affil-num=1
en-affil=
kn-affil=Department of Tumor Virology, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences
affil-num=2
en-affil=
kn-affil=Department of Tumor Virology, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences
affil-num=3
en-affil=
kn-affil=Department of Tumor Virology, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences
affil-num=4
en-affil=
kn-affil=Department of Tumor Virology, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences
affil-num=5
en-affil=
kn-affil=Research Center for Hepatitis and Immunology, National Center for Global Health and Medicine
affil-num=6
en-affil=
kn-affil=Research Center for Hepatitis and Immunology, National Center for Global Health and Medicine
affil-num=7
en-affil=
kn-affil=Department of Tumor Virology, Okayama University Graduate School of Medicine, Dentistry
affil-num=8
en-affil=
kn-affil=Department of Tumor Virology, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences
en-keyword=Antiviral response
kn-keyword=Antiviral response
en-keyword=hepatitis B virus
kn-keyword=hepatitis B virus
en-keyword=innate immune response
kn-keyword=innate immune response
en-keyword=cGAS-STING signaling pathway
kn-keyword=cGAS-STING signaling pathway
en-keyword=viral assembly
kn-keyword=viral assembly
END
start-ver=1.4
cd-journal=joma
no-vol=69
cd-vols=
no-issue=2
article-no=
start-page=71
end-page=78
dt-received=
dt-revised=
dt-accepted=
dt-pub-year=2015
dt-pub=201504
dt-online=
en-article=
kn-article=
en-subject=
kn-subject=
en-title=
kn-title=Annexin A1 Negatively Regulates Viral RNA Replication of Hepatitis C Virus
en-subtitle=
kn-subtitle=
en-abstract=
kn-abstract=Persistent infection with hepatitis C virus (HCV) often causes chronic hepatitis, and then shows a high rate of progression to liver cirrhosis and hepatocellular carcinoma. To clarify the mechanism of the persistent HCV infection is considered to be important for the discovery of new target(s) for the development of anti-HCV strategies. In the present study, we found that the expression level of annexin A1 (ANXA1) in human hepatoma cell line Li23-derived D7 cells was remarkably lower than that in parental Li23 cells, whereas the susceptibility of D7 cells to HCV infection was much higher than that of Li23 cells. Therefore, we hypothesized that ANXA1 negatively regulates persistent HCV infection through the inhibition of viral RNA replication. The results revealed that HCV production was significantly inhibited without a concomitant reduction in the amount of lipid droplets in the D7 cells stably expressing exogenous ANXA1. Further, we demonstrated that ANXA1 negatively regulated the step of viral RNA replication rather than that of viral entry in human hepatocytes. These results suggest that ANXA1 would be a novel target for the development of anti-HCV strategies.
en-copyright=
kn-copyright=
en-aut-name=HiramotoHiroki
en-aut-sei=Hiramoto
en-aut-mei=Hiroki
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=1
ORCID=
en-aut-name=DansakoHiromichi
en-aut-sei=Dansako
en-aut-mei=Hiromichi
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=2
ORCID=
en-aut-name=TakedaMidori
en-aut-sei=Takeda
en-aut-mei=Midori
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=3
ORCID=
en-aut-name=SatohShinya
en-aut-sei=Satoh
en-aut-mei=Shinya
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=4
ORCID=
en-aut-name=WakitaTakaji
en-aut-sei=Wakita
en-aut-mei=Takaji
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=5
ORCID=
en-aut-name=IkedaMasanori
en-aut-sei=Ikeda
en-aut-mei=Masanori
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=6
ORCID=
en-aut-name=KatoNobuyuki
en-aut-sei=Kato
en-aut-mei=Nobuyuki
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=7
ORCID=
affil-num=1
en-affil=
kn-affil=Department of Tumor Virology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences
affil-num=2
en-affil=
kn-affil=Department of Tumor Virology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences
affil-num=3
en-affil=
kn-affil=Department of Tumor Virology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences
affil-num=4
en-affil=
kn-affil=Department of Tumor Virology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences
affil-num=5
en-affil=
kn-affil=Department of Virology II, National Institute of Infectious Disease
affil-num=6
en-affil=
kn-affil=Department of Tumor Virology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences
affil-num=7
en-affil=
kn-affil=Department of Tumor Virology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences
en-keyword=HCV
kn-keyword=HCV
en-keyword=annexin A1
kn-keyword=annexin A1
en-keyword=Li23 cell line
kn-keyword=Li23 cell line
en-keyword=Li23-derived D7 cells
kn-keyword=Li23-derived D7 cells
en-keyword=HCV-JFH-1
kn-keyword=HCV-JFH-1
END
start-ver=1.4
cd-journal=joma
no-vol=430
cd-vols=
no-issue=2
article-no=
start-page=592
end-page=597
dt-received=
dt-revised=
dt-accepted=
dt-pub-year=2013
dt-pub=20130111
dt-online=
en-article=
kn-article=
en-subject=
kn-subject=
en-title=
kn-title=PML tumor suppressor protein is required for HCV production
en-subtitle=
kn-subtitle=
en-abstract=
kn-abstract=PML tumor suppressor protein, which forms discrete nuclear structures termed PML-nuclear bodies, has been associated with several cellular functions, including cell proliferation, apoptosis and antiviral defense. Recently, it was reported that the HCV core protein colocalizes with PML in PML-NBs and abrogates the PML function through interaction with PML. However, role(s) of PML in HCV life cycle is unknown. To test whether or not PML affects HCV life cycle, we examined the level of secreted HCV core and the infectivity of HCV in the culture supernatants as well as the level of HCV RNA in HuH-7-derived RSc cells, in which HCV-JFH1 can infect and efficiently replicate, stably expressing short hairpin RNA targeted to PML. In this context, the level of secreted HCV core and the infectivity in the supernatants from PML knockdown cells was remarkably reduced, whereas the level of HCV RNA in the PML knockdown cells was not significantly affected in spite of very effective knockdown of PML. In fact, we showed that PML is unrelated to HCV RNA replication using the subgenomic HCV-JFH1 replicon RNA, JRN/3-5B. Furthermore, the infectivity of HCV-like particle in the culture supernatants was significantly reduced in PML knockdown JRN/3-5B cells expressing core to NS2 coding region of HCV-JFH1 genome using the trans-packaging system. Finally, we also demonstrated that INI1 and DDX5, the PML-related proteins, are involved in HCV production. Taken together, these findings suggest that PML is required for HCV production.
en-copyright=
kn-copyright=
en-aut-name=KurokiMisao
en-aut-sei=Kuroki
en-aut-mei=Misao
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=1
ORCID=
en-aut-name=AriumiYasuo
en-aut-sei=Ariumi
en-aut-mei=Yasuo
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=2
ORCID=
en-aut-name=HijikataMakoto
en-aut-sei=Hijikata
en-aut-mei=Makoto
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=3
ORCID=
en-aut-name=IkedaMasanori
en-aut-sei=Ikeda
en-aut-mei=Masanori
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=4
ORCID=
en-aut-name=DansakoHiromichi
en-aut-sei=Dansako
en-aut-mei=Hiromichi
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=5
ORCID=
en-aut-name=WakitaTakaji
en-aut-sei=Wakita
en-aut-mei=Takaji
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=6
ORCID=
en-aut-name=ShimotohnoKunitada
en-aut-sei=Shimotohno
en-aut-mei=Kunitada
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=7
ORCID=
en-aut-name=KatoNobuyuki
en-aut-sei=Kato
en-aut-mei=Nobuyuki
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=8
ORCID=
affil-num=1
en-affil=
kn-affil=Okayama Univ, Dept Tumor Virol, Sch Med Dent & Pharmaceut Sci
affil-num=2
en-affil=
kn-affil=Okayama Univ, Dept Tumor Virol, Sch Med Dent & Pharmaceut Sci
affil-num=3
en-affil=
kn-affil=Kyoto Univ, Dept Viral Oncol, Inst Virus Res
affil-num=4
en-affil=
kn-affil=Okayama Univ, Dept Tumor Virol, Sch Med Dent & Pharmaceut Sci
affil-num=5
en-affil=
kn-affil=Okayama Univ, Dept Tumor Virol, Sch Med Dent & Pharmaceut Sci
affil-num=6
en-affil=
kn-affil=Natl Inst Infect Dis, Dept Virol 2
affil-num=7
en-affil=
kn-affil=Natl Ctr Global Hlth & Med, Res Ctr Hepatitis & Immunol
affil-num=8
en-affil=
kn-affil=Okayama Univ, Dept Tumor Virol, Sch Med Dent & Pharmaceut Sci
en-keyword=Hepatitis C virus
kn-keyword=Hepatitis C virus
en-keyword=PML
kn-keyword=PML
en-keyword=INI1
kn-keyword=INI1
en-keyword=DDX5
kn-keyword=DDX5
en-keyword=Tumor suppressor
kn-keyword=Tumor suppressor
en-keyword=Lipid droplet
kn-keyword=Lipid droplet
END
start-ver=1.4
cd-journal=joma
no-vol=66
cd-vols=
no-issue=6
article-no=
start-page=461
end-page=468
dt-received=
dt-revised=
dt-accepted=
dt-pub-year=2012
dt-pub=201212
dt-online=
en-article=
kn-article=
en-subject=
kn-subject=
en-title=
kn-title=Eradication of Hepatitis C Virus Subgenomic Replicon by Interferon Results in Aberrant Retinol-Related Protein Expression
en-subtitle=
kn-subtitle=
en-abstract=
kn-abstract=Hepatitis C virus (HCV) infection induces several changes in hepatocytes, such as oxidative stress, steatosis, and hepatocarcinogenesis. Although considerable progress has been made during recent years, the mechanisms underlying these functions remain unclear. We employed proteomic techniques in HCV replicon-harboring cells to determine the effects of HCV replication on host-cell protein expression. We examined two-dimensional electrophoresis (2-DE) and mass spectrometry to compare and identify differentially expressed proteins between HCV subgenomic replicon-harboring cells and their “cured” cells. One of the identified proteins was confirmed using enzyme-linked immunosorbent assay (ELISA) and Western blot analysis. Full-length HCV genome RNA replicating and cured cells were also assessed using ELISA. Replicon-harboring cells showed higher expression of retinal dehydrogenase 1 (RALDH-1), which converts retinol to retinoic acid, and the cured cells showed higher expression of retinol-binding protein (RBP), which transports retinol from the liver to target tissues. The alteration in RBP expression was also confirmed by ELISA and Western blot analysis. We conclude that protein expression profiling demonstrated that HCV replicon eradication affected retinol-related protein expression.
en-copyright=
kn-copyright=
en-aut-name=KoikeKazuko
en-aut-sei=Koike
en-aut-mei=Kazuko
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=1
ORCID=
en-aut-name=TakakiAkinobu
en-aut-sei=Takaki
en-aut-mei=Akinobu
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=2
ORCID=
en-aut-name=KatoNobuyuki
en-aut-sei=Kato
en-aut-mei=Nobuyuki
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=3
ORCID=
en-aut-name=OuchidaMamoru
en-aut-sei=Ouchida
en-aut-mei=Mamoru
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=4
ORCID=
en-aut-name=KanzakiHirotaka
en-aut-sei=Kanzaki
en-aut-mei=Hirotaka
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=5
ORCID=
en-aut-name=YasunakaTetsuya
en-aut-sei=Yasunaka
en-aut-mei=Tetsuya
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=6
ORCID=
en-aut-name=ShirahaHidenori
en-aut-sei=Shiraha
en-aut-mei=Hidenori
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=7
ORCID=
en-aut-name=MiyakeYasuhiro
en-aut-sei=Miyake
en-aut-mei=Yasuhiro
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=8
ORCID=
en-aut-name=YamamotoKazuhide
en-aut-sei=Yamamoto
en-aut-mei=Kazuhide
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=9
ORCID=
affil-num=1
en-affil=
kn-affil=
affil-num=2
en-affil=
kn-affil=
affil-num=3
en-affil=
kn-affil=
affil-num=4
en-affil=
kn-affil=
affil-num=5
en-affil=
kn-affil=
affil-num=6
en-affil=
kn-affil=
affil-num=7
en-affil=
kn-affil=
affil-num=8
en-affil=
kn-affil=
affil-num=9
en-affil=
kn-affil=
en-keyword=hepatitis C virus
kn-keyword=hepatitis C virus
en-keyword=retinol-binding protein
kn-keyword=retinol-binding protein
END
start-ver=1.4
cd-journal=joma
no-vol=409
cd-vols=
no-issue=4
article-no=
start-page=663
end-page=668
dt-received=
dt-revised=
dt-accepted=
dt-pub-year=2011
dt-pub=20110617
dt-online=
en-article=
kn-article=
en-subject=
kn-subject=
en-title=
kn-title=Plural assay systems derived from different cell lines and hepatitis C virus strains are required for the objective evaluation of anti-hepatitis C virus reagents
en-subtitle=
kn-subtitle=
en-abstract=
kn-abstract=Persistent hepatitis C virus (HCV) infection causes chronic liver diseases and is a global health problem. HuH-7 hepatoma-derived cells are widely used as the only cell-based HCV replication system for HCV research, including drug assays. Recently, using different hepatoma Li23-derived cells, we developed an HCV drug assay system (ORL8), in which the genome-length HCV RNA (O strain of genotype 1b) encoding renilla luciferase replicates efficiently. In this study, using the HuH-7-derived OR6 assay system that we developed previously and the ORL8 assay system, we evaluated 26 anti-HCV reagents, which other groups had reported as anti-HCV candidates using HuH-7-derived assay systems other than ORB. The results revealed that more than half of the reagents showed different anti-HCV activities from those in the previous studies, and that anti-HCV activities evaluated by the ORB and ORL8 assays were also frequently different. In further evaluation using the HuH-7-derived AH1R assay system, which was developed using the AH1 strain of genotype 1b, several reagents showed different anti-HCV activities in comparison with those evaluated by the OR6 and ORL8 assays. These results suggest that the different activities of anti-HCV reagents are caused by the differences in cell lines or HCV strains used for the development of assay systems. Therefore, we conclude that plural HCV assay systems developed using different cell lines or HCV strains are required for the objective evaluation of anti-HCV reagents.
en-copyright=
kn-copyright=
en-aut-name=UedaYouki
en-aut-sei=Ueda
en-aut-mei=Youki
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=1
ORCID=
en-aut-name=MoriKyoko
en-aut-sei=Mori
en-aut-mei=Kyoko
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=2
ORCID=
en-aut-name=AriumiYasuo
en-aut-sei=Ariumi
en-aut-mei=Yasuo
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=3
ORCID=
en-aut-name=IkedaMasanori
en-aut-sei=Ikeda
en-aut-mei=Masanori
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=4
ORCID=
en-aut-name=KatoNobuyuki
en-aut-sei=Kato
en-aut-mei=Nobuyuki
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=5
ORCID=
affil-num=1
en-affil=
kn-affil=Department of Tumor Virology, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences
affil-num=2
en-affil=
kn-affil=Department of Tumor Virology, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences
affil-num=3
en-affil=
kn-affil=Department of Tumor Virology, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences
affil-num=4
en-affil=
kn-affil=Department of Tumor Virology, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences
affil-num=5
en-affil=
kn-affil=Department of Tumor Virology, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences
en-keyword=HCV
kn-keyword=HCV
en-keyword=HCV RNA replication system
kn-keyword=HCV RNA replication system
en-keyword=Li23 cells
kn-keyword=Li23 cells
en-keyword=Reporter assay for anti-HCV reagents
kn-keyword=Reporter assay for anti-HCV reagents
END
start-ver=1.4
cd-journal=joma
no-vol=54
cd-vols=
no-issue=6
article-no=
start-page=253
end-page=257
dt-received=
dt-revised=
dt-accepted=
dt-pub-year=2000
dt-pub=200012
dt-online=
en-article=
kn-article=
en-subject=
kn-subject=
en-title=
kn-title=A reliable internally controlled RT-nested PCR method for the detection of hepatitis C virus RNA.
en-subtitle=
kn-subtitle=
en-abstract=
kn-abstract=We have developed a reliable internally controlled RT-nested PCR method for the detection of hepatitis C virus (HCV) RNA using in vitro synthesized Renilla luciferase (Rluc) RNA as an internal control. Using this method, the 5'-noncoding region of HCV RNA (144 nucleotides) and Rluc RNA (276 nucleotides) were efficiently amplified in a single tube, and the sensitivity and specificity of this method were comparable to standard RT-nested PCR. This method was successfully performed on RNA specimens obtained from in vitro HCV-infected human hepatocyte PH5CH8 cells, which support HCV replication. In addition, we demonstrated that this method was useful for the evaluation of antiviral reagents by confirming the anti-HCV activity of bovine lactoferrin, which we previously found to be a new inhibitor of HCV infection. Therefore, this method may be useful for the studies of not only HCV but also of other viruses.
en-copyright=
kn-copyright=
en-aut-name=NozakiAkito
en-aut-sei=Nozaki
en-aut-mei=Akito
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=1
ORCID=
en-aut-name=NaganumaAtsushi
en-aut-sei=Naganuma
en-aut-mei=Atsushi
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=2
ORCID=
en-aut-name=NakamuraTakashi
en-aut-sei=Nakamura
en-aut-mei=Takashi
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=3
ORCID=
en-aut-name=TanakaKatsuaki
en-aut-sei=Tanaka
en-aut-mei=Katsuaki
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=4
ORCID=
en-aut-name=SekiharaHisahiko
en-aut-sei=Sekihara
en-aut-mei=Hisahiko
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=5
ORCID=
en-aut-name=KatoNobuyuki
en-aut-sei=Kato
en-aut-mei=Nobuyuki
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=6
ORCID=
affil-num=1
en-affil=
kn-affil=Yokohama City University
affil-num=2
en-affil=
kn-affil=Okayama University
affil-num=3
en-affil=
kn-affil=Okayama University
affil-num=4
en-affil=
kn-affil=Yokohama City University
affil-num=5
en-affil=
kn-affil=Yokohama City University
affil-num=6
en-affil=
kn-affil=Okayama University
en-keyword=Hepatitis C virus
kn-keyword=Hepatitis C virus
en-keyword= Reverse transcriptionnested PCR (RT-nested PCR)
kn-keyword= Reverse transcriptionnested PCR (RT-nested PCR)
en-keyword=internal control
kn-keyword=internal control
END
start-ver=1.4
cd-journal=joma
no-vol=55
cd-vols=
no-issue=3
article-no=
start-page=133
end-page=159
dt-received=
dt-revised=
dt-accepted=
dt-pub-year=2001
dt-pub=200106
dt-online=
en-article=
kn-article=
en-subject=
kn-subject=
en-title=
kn-title=Molecular virology of hepatitis C virus.
en-subtitle=
kn-subtitle=
en-abstract=
kn-abstract=Hepatitis C virus (HCV), discovered in 1989, is the major causative agent of parenteral non-A, non-B hepatitis worldwide. Following the development of a method of diagnosing HCV infection, it became apparent that HCV frequently causes chronic hepatitis. Persistent infection with HCV is implicated in liver cirrhosis and hepatocellular carcinoma. Current worldwide estimations suggest that more than 170 million people have been infected with HCV, an enveloped positive single-stranded RNA (9.6-kilobases) virus belonging to the Flaviviridae. The HCV genome shows remarkable sequence variation, especially in the hypervariable region 1 of the E2 protein-encoding region, and globally, HCV appears to be distributed with more than 30 genotypes. Complicated "quasispecies" and frequent mutations of viral genomes have also emerged. The HCV genome encodes a large polyprotein precursor of about 3,000 amino acid residues, and this precursor protein is cleaved by the host and viral proteinases to generate at least 10 proteins in the following order: NH2-core-envelope (E1)-E2-p7-nonstructural protein 2 (NS2)-NS3-NS4A-NS4B-NS5A-NS5B-COOH. These viral proteins not only function in viral replication but also affect a variety of cellular functions. Although several explanations have been proposed, the mechanisms of HCV infection and replication in targeted cells, the mechanism of persistent viral infection, and the pathogenesis of hepatic diseases (hepatitis or hepatocellular carcinoma) are all poorly understood. A major reason why these mechanisms remain unclear is the lack of a good experimental HCV replication system. Although several classical trials using cultured cells have been reported, several new, more promising experimental strategies (generations of infectious cDNA clone, replicon, animal models, etc.) are currently being designed and tested, in order to resolve these problems. In addition, new therapies for chronic hepatitis have also been developed. The enormous body of information collected thus far in the field of HCV research is summarized below, and an overview of the current status of HCV molecular virology of HCV is provided.</P>
en-copyright=
kn-copyright=
en-aut-name=KatoNobuyuki
en-aut-sei=Kato
en-aut-mei=Nobuyuki
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=1
ORCID=
affil-num=1
en-affil=
kn-affil=Okayama University
END
start-ver=1.4
cd-journal=joma
no-vol=56
cd-vols=
no-issue=3
article-no=
start-page=141
end-page=147
dt-received=
dt-revised=
dt-accepted=
dt-pub-year=2002
dt-pub=200206
dt-online=
en-article=
kn-article=
en-subject=
kn-subject=
en-title=
kn-title=Hepatitis C virus quasispecies in cancerous and noncancerous hepatic lesions: the core protein-encoding region.
en-subtitle=
kn-subtitle=
en-abstract=
kn-abstract=We have shown that highly proofreading DNA polymerase is required for the polymerase chain reaction in the genetic analysis of hepatitis C virus (HCV). To clarify the status of HCV quasispecies in hepatic tissue using proofreading DNA polymerase, we performed a genetic analysis of the HCV core protein-encoding region in cancerous and noncancerous lesions derived from 4 patients with hepatocellular carcinoma. In contrast to the previously published data, we observed neither deletions nor stop codons in the analyzed region and no significant difference in the complexity of HCV quasispecies between cancerous and noncancerous lesions. This result suggests that the HCV core gene is never structurally defective in hepatic tissues, including cancerous lesions. However, in 3 of the patients, the consensus HCV species differed between cancerous and noncancerous lesions, suggesting that the predominant replicating HCV species differs between these 2 types of lesions. Moreover, during the course of the study, we obtained several interesting variants possessing a substitution at codon 9 of the core gene, whose substitution has been shown to induce the production of the F protein synthesized by a - 2/+1 ribosomal frameshift.
en-copyright=
kn-copyright=
en-aut-name=AlamShahjalal S.
en-aut-sei=Alam
en-aut-mei=Shahjalal S.
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=1
ORCID=
en-aut-name=NakamuraTakashi
en-aut-sei=Nakamura
en-aut-mei=Takashi
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=2
ORCID=
en-aut-name=NaganumaAtsushi
en-aut-sei=Naganuma
en-aut-mei=Atsushi
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=3
ORCID=
en-aut-name=NozakiAkito
en-aut-sei=Nozaki
en-aut-mei=Akito
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=4
ORCID=
en-aut-name=NousoKazuhiro
en-aut-sei=Nouso
en-aut-mei=Kazuhiro
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=5
ORCID=
en-aut-name=ShimomuraHiroyuki
en-aut-sei=Shimomura
en-aut-mei=Hiroyuki
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=6
ORCID=
en-aut-name=KatoNobuyuki
en-aut-sei=Kato
en-aut-mei=Nobuyuki
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=7
ORCID=
affil-num=1
en-affil=
kn-affil=Okayama University
affil-num=2
en-affil=
kn-affil=Okayama University
affil-num=3
en-affil=
kn-affil=Okayama University
affil-num=4
en-affil=
kn-affil=Okayama University
affil-num=5
en-affil=
kn-affil=Okayama University
affil-num=6
en-affil=
kn-affil=Okayama University
affil-num=7
en-affil=
kn-affil=Okayama University
en-keyword=hepatitis C virus
kn-keyword=hepatitis C virus
en-keyword=core gene
kn-keyword=core gene
en-keyword=hepatocellular carcinoma
kn-keyword=hepatocellular carcinoma
en-keyword=quasispecies
kn-keyword=quasispecies
en-keyword=proofreading DNA polymerase
kn-keyword=proofreading DNA polymerase
END
start-ver=1.4
cd-journal=joma
no-vol=56
cd-vols=
no-issue=2
article-no=
start-page=107
end-page=110
dt-received=
dt-revised=
dt-accepted=
dt-pub-year=2002
dt-pub=200204
dt-online=
en-article=
kn-article=
en-subject=
kn-subject=
en-title=
kn-title=Quantitative method of intracellular hepatitis C virus RNA using LightCycler PCR.
en-subtitle=
kn-subtitle=
en-abstract=
kn-abstract=Based on recent LightCycler techniques developed for the quantitation of serum HCV RNA, we have developed a quantitative method for the intracellular hepatitis C virus (HCV) RNA using LightCycler PCR. A simple real-time PCR assay, based on the SYBR Green I dye and LightCycler fluorimeter and with no probe requirement, is described. In the presence of 0.5 microg of cellular RNA, it was demonstrated that as few as 25 copies of HCV RNA could be specifically detected with a set of primers that amplify a 144-base pair sequence unique to the 5'-noncoding region of HCV RNA. We demonstrated that this method was useful for the evaluation of antiviral reagents using HCV-infected human cultured cells.
en-copyright=
kn-copyright=
en-aut-name=NozakiAkito
en-aut-sei=Nozaki
en-aut-mei=Akito
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=1
ORCID=
en-aut-name=KatoNobuyuki
en-aut-sei=Kato
en-aut-mei=Nobuyuki
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=2
ORCID=
affil-num=1
en-affil=
kn-affil=Okayama University
affil-num=2
en-affil=
kn-affil=Okayama University
en-keyword=hepatitis C virus
kn-keyword=hepatitis C virus
en-keyword=real-time PCR
kn-keyword=real-time PCR
en-keyword=LightCycler
kn-keyword=LightCycler
END
start-ver=1.4
cd-journal=joma
no-vol=122
cd-vols=
no-issue=1
article-no=
start-page=9
end-page=16
dt-received=
dt-revised=
dt-accepted=
dt-pub-year=2010
dt-pub=20100401
dt-online=
en-article=
kn-article=
en-subject=
kn-subject=
en-title=The DNA damage sensors ataxia-telangiectasia mutated kinase and checkpoint kinase 2 are required for hepatitis C virus RNA replication
kn-title=DNA 損傷センサーATMキナーゼとChk2はＣ型肝炎ウイルスのRNA複製に必要である
en-subtitle=
kn-subtitle=
en-abstract=
kn-abstract=
en-copyright=
kn-copyright=
en-aut-name=AriumiYasuo
en-aut-sei=Ariumi
en-aut-mei=Yasuo
kn-aut-name=有海康雄
kn-aut-sei=有海
kn-aut-mei=康雄
aut-affil-num=1
ORCID=
en-aut-name=KurokiMisao
en-aut-sei=Kuroki
en-aut-mei=Misao
kn-aut-name=黒木美沙緒
kn-aut-sei=黒木
kn-aut-mei=美沙緒
aut-affil-num=2
ORCID=
en-aut-name=DansakoHiromichi
en-aut-sei=Dansako
en-aut-mei=Hiromichi
kn-aut-name=團迫浩方
kn-aut-sei=團迫
kn-aut-mei=浩方
aut-affil-num=3
ORCID=
en-aut-name=AbeKenichi
en-aut-sei=Abe
en-aut-mei=Kenichi
kn-aut-name=阿部健一
kn-aut-sei=阿部
kn-aut-mei=健一
aut-affil-num=4
ORCID=
en-aut-name=IkedaMasanori
en-aut-sei=Ikeda
en-aut-mei=Masanori
kn-aut-name=池田正徳
kn-aut-sei=池田
kn-aut-mei=正徳
aut-affil-num=5
ORCID=
en-aut-name=WakitaTakaji
en-aut-sei=Wakita
en-aut-mei=Takaji
kn-aut-name=脇田隆字
kn-aut-sei=脇田
kn-aut-mei=隆字
aut-affil-num=6
ORCID=
en-aut-name=KatoNobuyuki
en-aut-sei=Kato
en-aut-mei=Nobuyuki
kn-aut-name=加藤宣之
kn-aut-sei=加藤
kn-aut-mei=宣之
aut-affil-num=7
ORCID=
affil-num=1
en-affil=
kn-affil=岡山大学大学院医歯薬学総合研究科 腫瘍ウイルス学
affil-num=2
en-affil=
kn-affil=岡山大学大学院医歯薬学総合研究科 腫瘍ウイルス学
affil-num=3
en-affil=
kn-affil=岡山大学大学院医歯薬学総合研究科 腫瘍ウイルス学
affil-num=4
en-affil=
kn-affil=岡山大学大学院医歯薬学総合研究科 腫瘍ウイルス学
affil-num=5
en-affil=
kn-affil=岡山大学大学院医歯薬学総合研究科 腫瘍ウイルス学
affil-num=6
en-affil=
kn-affil=国立感染症研究所 ウイルス第二部
affil-num=7
en-affil=
kn-affil=岡山大学大学院医歯薬学総合研究科 腫瘍ウイルス学
en-keyword=HCV
kn-keyword=HCV
en-keyword=ATM
kn-keyword=ATM
en-keyword=Chk2
kn-keyword=Chk2
en-keyword=宿主因子
kn-keyword=宿主因子
en-keyword=DNA 損傷センサー
kn-keyword=DNA 損傷センサー
END
start-ver=1.4
cd-journal=joma
no-vol=113
cd-vols=
no-issue=1
article-no=
start-page=101
end-page=106
dt-received=
dt-revised=
dt-accepted=
dt-pub-year=2001
dt-pub=20010428
dt-online=
en-article=
kn-article=
en-subject=
kn-subject=
en-title=
kn-title=感染症と癌
en-subtitle=
kn-subtitle=
en-abstract=
kn-abstract=
en-copyright=
kn-copyright=
en-aut-name=
en-aut-sei=
en-aut-mei=
kn-aut-name=加藤宣之
kn-aut-sei=加藤
kn-aut-mei=宣之
aut-affil-num=1
ORCID=
en-aut-name=
en-aut-sei=
en-aut-mei=
kn-aut-name=小熊惠二
kn-aut-sei=小熊
kn-aut-mei=惠二
aut-affil-num=2
ORCID=
affil-num=1
en-affil=
kn-affil=岡山大学医学部病態分子生物学講座
affil-num=2
en-affil=
kn-affil=岡山大学医学部細菌学講座
en-keyword=C型肝炎ウィルス
kn-keyword=C型肝炎ウィルス
en-keyword=EBウィルス
kn-keyword=EBウィルス
en-keyword=Helicobacter pylori
kn-keyword=Helicobacter pylori
END