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Although viruses infect various organs and are associated with diseases, there may be many unidentified pathogenic viruses. The recent development of next-generation sequencing technologies has facilitated the establishment of an environmental viral metagenomic approach targeting the intracellular viral genome. However, an efficient method for the detection of a viral genome derived from an RNA virus in animal or human samples has not been established. Here, we established a method for the efficient detection of RNA viruses in human clinical samples. We then tested the efficiency of the method compared to other conventional methods by using tissue samples collected from 57 recipients of living donor liver transplantations performed between June 2017 and February 2019 at Kyushu University Hospital. The viral read ratio in human clinical samples was higher by the new method than by the other conventional methods. In addition, the new method correctly identified viral RNA from liver tissues infected with hepatitis C virus. This new technique will be an effective tool for intracellular RNA virus surveillance in human clinical samples and may be useful for the detection of new RNA viruses associated with diseases.
Takuma Izumi; Yuhei Morioka; Syun-Ichi Urayama; Daisuke Motooka; Tomokazu Tamura; Takahiro Kawagishi; Yuta Kanai; Takeshi Kobayashi; Chikako Ono; Akinari Morinaga; Takahiro Tomiyama; Norifumi Iseda; Yukiko Kosai; Shoichi Inokuchi; Shota Nakamura; Tomohisa Tanaka; Kohji Moriishi; Hiroaki Kariwa; Tomoharu Yoshizumi; Masaki Mori; Yoshiharu Matsuura; Takasuke Fukuhara. DsRNA Sequencing for RNA Virus Surveillance Using Human Clinical Samples. Viruses 2021, 13, 1310 .
AMA StyleTakuma Izumi, Yuhei Morioka, Syun-Ichi Urayama, Daisuke Motooka, Tomokazu Tamura, Takahiro Kawagishi, Yuta Kanai, Takeshi Kobayashi, Chikako Ono, Akinari Morinaga, Takahiro Tomiyama, Norifumi Iseda, Yukiko Kosai, Shoichi Inokuchi, Shota Nakamura, Tomohisa Tanaka, Kohji Moriishi, Hiroaki Kariwa, Tomoharu Yoshizumi, Masaki Mori, Yoshiharu Matsuura, Takasuke Fukuhara. DsRNA Sequencing for RNA Virus Surveillance Using Human Clinical Samples. Viruses. 2021; 13 (7):1310.
Chicago/Turabian StyleTakuma Izumi; Yuhei Morioka; Syun-Ichi Urayama; Daisuke Motooka; Tomokazu Tamura; Takahiro Kawagishi; Yuta Kanai; Takeshi Kobayashi; Chikako Ono; Akinari Morinaga; Takahiro Tomiyama; Norifumi Iseda; Yukiko Kosai; Shoichi Inokuchi; Shota Nakamura; Tomohisa Tanaka; Kohji Moriishi; Hiroaki Kariwa; Tomoharu Yoshizumi; Masaki Mori; Yoshiharu Matsuura; Takasuke Fukuhara. 2021. "DsRNA Sequencing for RNA Virus Surveillance Using Human Clinical Samples." Viruses 13, no. 7: 1310.
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the causative agent of coronavirus disease 2019 (COVID-19), employs host-cell angiotensin-converting enzyme 2 (ACE2) for cell entry. Genetic analyses of ACE2 have identified several single-nucleotide polymorphisms (SNPs) specific to different human populations. Molecular dynamics simulations have indicated that several of these SNPs could affect interactions between SARS-CoV-2 and ACE2, thereby providing a partial explanation for the regional differences observed in SARS-CoV-2 infectivity and severity. However, the significance of population-specific ACE2 SNPs in SARS-CoV-2 infectivity is unknown, as no in vitro validation studies have been performed. Here, we analyzed the impact of eight SNPs found in specific populations on receptor binding and cell entry in vitro. Except for a SNP causing a nonsense mutation that reduced ACE2 expression, none of the selected SNPs markedly altered the interaction between ACE2 and the SARS-CoV-2 spike protein (SARS-2-S), which is responsible for receptor recognition and cell entry, or the efficiency of viral cell entry mediated by SARS-2-S. Our findings indicate that ACE2 polymorphisms have limited impact on the ACE2-dependent cell entry of SARS-CoV-2 and underscore the importance of future studies on the involvement of population-specific SNPs of other host genes in susceptibility toward SARS-CoV-2 infection.
Mei Hashizume; Gabriel Gonzalez; Chikako Ono; Ayako Takashima; Masaharu Iwasaki. Population-Specific ACE2 Single-Nucleotide Polymorphisms Have Limited Impact on SARS-CoV-2 Infectivity In Vitro. Viruses 2021, 13, 67 .
AMA StyleMei Hashizume, Gabriel Gonzalez, Chikako Ono, Ayako Takashima, Masaharu Iwasaki. Population-Specific ACE2 Single-Nucleotide Polymorphisms Have Limited Impact on SARS-CoV-2 Infectivity In Vitro. Viruses. 2021; 13 (1):67.
Chicago/Turabian StyleMei Hashizume; Gabriel Gonzalez; Chikako Ono; Ayako Takashima; Masaharu Iwasaki. 2021. "Population-Specific ACE2 Single-Nucleotide Polymorphisms Have Limited Impact on SARS-CoV-2 Infectivity In Vitro." Viruses 13, no. 1: 67.
The discovery of novel antivirals to treat hepatitis B virus (HBV) infection is urgently needed, as the currently available drugs mainly target viral proteins at replication step, whereas host factors also play significant roles in HBV infection. Although numerous studies have reported candidate drugs for HBV treatment, there remains a need to find a new drug that may target other steps of the HBV life cycle. In this study, by drug screening of a 533 G-protein-coupled receptors (GPCRs)-associated compound library, we identified ponesimod, a selective agonist of sphingosine-1-phosphate receptor 1 (S1P1), as a drug candidate for the suppression of HBV infection. However, the anti-HBV effect of ponesimod is independent of S1P1 and other sphingosine-1-phosphate receptors (S1PRs). Treatment with ponesimod at an early step of infection but not at a post-entry step significantly reduced the HBV relaxed circular DNA (rcDNA) level in a dose-dependent manner. Ponesimod treatment did not inhibit attachment, binding, or internalization of HBV particles via endocytosis through an interaction with sodium taurocholate cotransporting polypeptide (NTCP) or epidermal growth factor receptor (EGFR). Importantly, during the transportation of HBV particles to the nucleus, co-localization of HBV with early endosomes but not with late endosomes and lysosomes was induced by the treatment with ponesimod, suggesting that ponesimod interferes with the conversion of early endosomes to late endosomes without significant damage to cellular growth. Conclusion: Ponesimod is a promising anti-HBV drug targeting the endosome maturation of HBV. This finding can be applied to the development of novel antivirals that target the trafficking pathway of HBV particles.
Yuzy Fauzyah; Chikako Ono; Shiho Torii; Itsuki Anzai; Rigel Suzuki; Takuma Izumi; Yuhei Morioka; Yusuke Maeda; Toru Okamoto; Takasuke Fukuhara; Yoshiharu Matsuura. Ponesimod suppresses hepatitis B virus infection by inhibiting endosome maturation. Antiviral Research 2020, 186, 104999 .
AMA StyleYuzy Fauzyah, Chikako Ono, Shiho Torii, Itsuki Anzai, Rigel Suzuki, Takuma Izumi, Yuhei Morioka, Yusuke Maeda, Toru Okamoto, Takasuke Fukuhara, Yoshiharu Matsuura. Ponesimod suppresses hepatitis B virus infection by inhibiting endosome maturation. Antiviral Research. 2020; 186 ():104999.
Chicago/Turabian StyleYuzy Fauzyah; Chikako Ono; Shiho Torii; Itsuki Anzai; Rigel Suzuki; Takuma Izumi; Yuhei Morioka; Yusuke Maeda; Toru Okamoto; Takasuke Fukuhara; Yoshiharu Matsuura. 2020. "Ponesimod suppresses hepatitis B virus infection by inhibiting endosome maturation." Antiviral Research 186, no. : 104999.
Summary Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has been identified as the causative agent of coronavirus disease 2019 (COVID-19). While the development of specific treatments and a vaccine is urgently needed, functional analyses of SARS-CoV-2 have been limited by the lack of convenient mutagenesis methods. In this study, we established a PCR-based, bacterium-free method to generate SARS-CoV-2 infectious clones. Recombinant SARS-CoV-2 could be rescued at high titer with high accuracy after assembling 10 SARS-CoV-2 cDNA fragments by circular polymerase extension reaction (CPER) and transfection of the resulting circular genome into susceptible cells. Notably, the construction of infectious clones for reporter viruses and mutant viruses could be completed in two simple steps: introduction of reporter genes or mutations into the desirable DNA fragments (~5,000 base pairs) by PCR and assembly of the DNA fragments by CPER. We hope that our reverse genetics system will contribute to the further understanding of SARS-CoV-2.
Shiho Torii; Chikako Ono; Rigel Suzuki; Yuhei Morioka; Itsuki Anzai; Yuzy Fauzyah; Yusuke Maeda; Wataru Kamitani; Takasuke Fukuhara; Yoshiharu Matsuura. Establishment of a reverse genetics system for SARS-CoV-2 using circular polymerase extension reaction. 2020, 1 .
AMA StyleShiho Torii, Chikako Ono, Rigel Suzuki, Yuhei Morioka, Itsuki Anzai, Yuzy Fauzyah, Yusuke Maeda, Wataru Kamitani, Takasuke Fukuhara, Yoshiharu Matsuura. Establishment of a reverse genetics system for SARS-CoV-2 using circular polymerase extension reaction. . 2020; ():1.
Chicago/Turabian StyleShiho Torii; Chikako Ono; Rigel Suzuki; Yuhei Morioka; Itsuki Anzai; Yuzy Fauzyah; Yusuke Maeda; Wataru Kamitani; Takasuke Fukuhara; Yoshiharu Matsuura. 2020. "Establishment of a reverse genetics system for SARS-CoV-2 using circular polymerase extension reaction." , no. : 1.
The aim of this study is to understand adaptive immunity to SARS-CoV-2 through the analysis of B cell epitope and neutralizing activity in coronavirus disease 2019 (COVID-19) patients. We obtained serum from thirteen COVID-19 patients. Most individuals revealed neutralizing activity against SARS-CoV-2 assessed by a pseudotype virus-neutralizing assay. The antibody production against the spike glycoprotein (S protein) or receptor-binding domain (RBD) of SARS-CoV-2 was elevated, with large individual differences, as assessed by ELISA. In the analysis of the predicted the linear B cell epitopes, two regions (671-690 aa. and 1146-1164 aa.), which were located in S1 and S2 but not in the RBD, were highly reactive with the sera from patients. In the further analysis of the B cell epitope within the S protein by utilizing a B cell epitope array, a hot spot in the N-terminal domain of the S protein but not the RBD was observed in individuals with neutralizing activity. Overall, the analysis of antibody production and B cell epitopes of the S protein from patient serum may provide a novel target for the vaccine development against SARS-CoV-2.
Shota Yoshida; Chikako Ono; Hiroki Hayashi; Satoshi Shiraishi; Kazunori Tomono; Hisashi Arase; Yoshiharu Matsuura; Hironori Nakagami. SARS-CoV-2-induced humoral immunity through B cell epitope analysis and neutralizing activity in COVID-19 infected individuals in Japan. 2020, 1 .
AMA StyleShota Yoshida, Chikako Ono, Hiroki Hayashi, Satoshi Shiraishi, Kazunori Tomono, Hisashi Arase, Yoshiharu Matsuura, Hironori Nakagami. SARS-CoV-2-induced humoral immunity through B cell epitope analysis and neutralizing activity in COVID-19 infected individuals in Japan. . 2020; ():1.
Chicago/Turabian StyleShota Yoshida; Chikako Ono; Hiroki Hayashi; Satoshi Shiraishi; Kazunori Tomono; Hisashi Arase; Yoshiharu Matsuura; Hironori Nakagami. 2020. "SARS-CoV-2-induced humoral immunity through B cell epitope analysis and neutralizing activity in COVID-19 infected individuals in Japan." , no. : 1.
Based on its ability to express high levels of protein, baculovirus has been widely used for recombinant protein production in insect cells for more than thirty years with continued technical improvements. In addition, baculovirus has been successfully applied for foreign gene delivery into mammalian cells without any viral replication. However, several CpG motifs are present throughout baculoviral DNA and induce an antiviral response in mammalian cells, resulting in the production of pro-inflammatory cytokines and type I interferon through a Toll-like receptor (TLR)-dependent or -independent signaling pathway, and ultimately limiting the efficiency of transgene expression. On the other hand, by taking advantage of this strong adjuvant activity, recombinant baculoviruses encoding neutralization epitopes can elicit protective immunity in mice. Moreover, immunodeficient cells, such as hepatitis C virus (HCV)- or human immunodeficiency virus (HIV)-infected cells, are more susceptible to baculovirus infection than normal cells and are selectively eliminated by the apoptosis-inducible recombinant baculovirus. Here, we summarize the application of baculovirus as a gene expression vector and the mechanism of the host innate immune response induced by baculovirus in mammalian cells. We also discuss the future prospects of baculovirus vectors.
Chikako Ono; Toru Okamoto; Takayuki Abe; Yoshiharu Matsuura. Baculovirus as a Tool for Gene Delivery and Gene Therapy. Viruses 2018, 10, 510 .
AMA StyleChikako Ono, Toru Okamoto, Takayuki Abe, Yoshiharu Matsuura. Baculovirus as a Tool for Gene Delivery and Gene Therapy. Viruses. 2018; 10 (9):510.
Chicago/Turabian StyleChikako Ono; Toru Okamoto; Takayuki Abe; Yoshiharu Matsuura. 2018. "Baculovirus as a Tool for Gene Delivery and Gene Therapy." Viruses 10, no. 9: 510.