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  • 1
    Online Resource
    Online Resource
    Establishment of a Virulent Full-Length cDNA Clone for Type I Feline Coronavirus Strain C3663
    American Society for Microbiology ; 2019
    In:  Journal of Virology Vol. 93, No. 21 ( 2019-11)
    Details
    In: Journal of Virology, American Society for Microbiology, Vol. 93, No. 21 ( 2019-11)
    Abstract: Feline infectious peritonitis (FIP) is one of the most important infectious diseases in cats and is caused by feline coronavirus (FCoV). Tissue culture-adapted type I FCoV shows reduced FIP induction in experimental infections, which complicates the understanding of FIP pathogenesis caused by type I FCoV. We previously found that the type I FCoV strain C3663 efficiently induces FIP in specific-pathogen-free cats through the naturally infectious route. In this study, we employed a bacterial artificial chromosome-based reverse genetics system to gain more insights into FIP caused by the C3633 strain. We successfully generated recombinant virus (rC3663) from Fcwf-4 cells transfected with infectious cDNA that showed growth kinetics similar to those shown by the parental virus. Next, we constructed a reporter C3663 virus carrying the nanoluciferase (Nluc) gene to measure viral replication with high sensitivity. The inhibitory effects of different compounds against rC3663-Nluc could be measured within 24 h postinfection. Furthermore, we found that A72 cells derived from canine fibroblasts permitted FCoV replication without apparent cytopathic effects. Thus, our reporter virus is useful for uncovering the infectivity of type I FCoV in different cell lines, including canine-derived cells. Surprisingly, we uncovered aberrant viral RNA transcription of rC3663 in A72 cells. Overall, we succeeded in obtaining infectious cDNA clones derived from type I FCoV that retained its virulence. Our recombinant FCoVs are powerful tools for increasing our understanding of the viral life cycle and pathogenesis of FIP-inducing type I FCoV. IMPORTANCE Feline coronavirus (FCoV) is one of the most significant coronaviruses, because this virus induces feline infectious peritonitis (FIP), which is a lethal disease in cats. Tissue culture-adapted type I FCoV often loses pathogenicity, which complicates research on type I FCoV-induced feline infectious peritonitis (FIP). Since we previously found that type I FCoV strain C3663 efficiently induces FIP in specific-pathogen-free cats, we established a reverse genetics system for the C3663 strain to obtain recombinant viruses in the present study. By using a reporter C3663 virus, we were able to examine the inhibitory effect of 68 compounds on C3663 replication in Fcwf-4 cells and infectivity in a canine-derived cell line. Interestingly, one canine cell line, A72, permitted FCoV replication but with low efficiency and aberrant viral gene expression.
    Type of Medium: Online Resource
    ISSN: 0022-538X , 1098-5514
    URL: Article
    DOI: 10.1128/JVI.01208-19
    Language: English
    Publisher: American Society for Microbiology
    Publication Date: 2019
    detail.hit.zdb_id: 1495529-5
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  • 2
    Online Resource
    Online Resource
    Broad-spectrum antiviral agents: secreted phospholipase A2 targets viral envelope lipid bilayers derived from the endoplasmic reticulum membrane
    Springer Science and Business Media LLC ; 2017
    In:  Scientific Reports Vol. 7, No. 1 ( 2017-11-21)
    Details
    In: Scientific Reports, Springer Science and Business Media LLC, Vol. 7, No. 1 ( 2017-11-21)
    Abstract: Hepatitis C virus (HCV), dengue virus (DENV) and Japanese encephalitis virus (JEV) belong to the family Flaviviridae . Their viral particles have the envelope composed of viral proteins and a lipid bilayer acquired from budding through the endoplasmic reticulum (ER). The phospholipid content of the ER membrane differs from that of the plasma membrane (PM). The phospholipase A 2 (PLA 2 ) superfamily consists of a large number of members that specifically catalyse the hydrolysis of phospholipids at a particular position. Here we show that the CM-II isoform of secreted PLA 2 obtained from Naja mossambica mossambica snake venom (CM-II-sPLA 2 ) possesses potent virucidal (neutralising) activity against HCV, DENV and JEV, with 50% inhibitory concentrations (IC 50 ) of 0.036, 0.31 and 1.34 ng/ml, respectively. In contrast, the IC 50 values of CM-II-sPLA 2 against viruses that bud through the PM (Sindbis virus, influenza virus and Sendai virus) or trans -Golgi network (TGN) (herpes simplex virus) were 〉 10,000 ng/ml. Moreover, the 50% cytotoxic (CC 50 ) and haemolytic (HC 50 ) concentrations of CM-II-sPLA 2 were 〉 10,000 ng/ml, implying that CM-II-sPLA 2 did not significantly damage the PM. These results suggest that CM-II-sPLA 2 and its derivatives are good candidates for the development of broad-spectrum antiviral drugs that target viral envelope lipid bilayers derived from the ER membrane.
    Type of Medium: Online Resource
    ISSN: 2045-2322
    URL: Article
    DOI: 10.1038/s41598-017-16130-w
    Language: English
    Publisher: Springer Science and Business Media LLC
    Publication Date: 2017
    detail.hit.zdb_id: 2615211-3
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  • 3
    Online Resource
    Online Resource
    MERS coronavirus nsp1 participates in an efficient propagation through a specific interaction with viral RNA
    Elsevier BV ; 2017
    In:  Virology Vol. 511 ( 2017-11), p. 95-105
    Details
    In: Virology, Elsevier BV, Vol. 511 ( 2017-11), p. 95-105
    Type of Medium: Online Resource
    ISSN: 0042-6822
    URL: Article
    DOI: 10.1016/j.virol.2017.08.026
    RVK:
    WA 15000
    Language: English
    Publisher: Elsevier BV
    Publication Date: 2017
    detail.hit.zdb_id: 1471925-3
    SSG: 12
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  • 4
    Online Resource
    Online Resource
    Two-amino acids change in the nsp4 of SARS coronavirus abolishes viral replication
    Elsevier BV ; 2017
    In:  Virology Vol. 510 ( 2017-10), p. 165-174
    Details
    In: Virology, Elsevier BV, Vol. 510 ( 2017-10), p. 165-174
    Type of Medium: Online Resource
    ISSN: 0042-6822
    URL: Article
    DOI: 10.1016/j.virol.2017.07.019
    RVK:
    WA 15000
    Language: English
    Publisher: Elsevier BV
    Publication Date: 2017
    detail.hit.zdb_id: 1471925-3
    SSG: 12
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  • 5
    Online Resource
    Online Resource
    Combining machine learning and nanopore construction creates an artificial intelligence nanopore for coronavirus detection
    Springer Science and Business Media LLC ; 2021
    In:  Nature Communications Vol. 12, No. 1 ( 2021-06-17)
    Details
    In: Nature Communications, Springer Science and Business Media LLC, Vol. 12, No. 1 ( 2021-06-17)
    Abstract: High-throughput, high-accuracy detection of emerging viruses allows for the control of disease outbreaks. Currently, reverse transcription-polymerase chain reaction (RT-PCR) is currently the most-widely used technology to diagnose the presence of SARS-CoV-2. However, RT-PCR requires the extraction of viral RNA from clinical specimens to obtain high sensitivity. Here, we report a method for detecting novel coronaviruses with high sensitivity by using nanopores together with artificial intelligence, a relatively simple procedure that does not require RNA extraction. Our final platform, which we call the artificially intelligent nanopore, consists of machine learning software on a server, a portable high-speed and high-precision current measuring instrument, and scalable, cost-effective semiconducting nanopore modules. We show that artificially intelligent nanopores are successful in accurately identifying four types of coronaviruses similar in size, HCoV-229E, SARS-CoV, MERS-CoV, and SARS-CoV-2. Detection of SARS-CoV-2 in saliva specimen is achieved with a sensitivity of 90% and specificity of 96% with a 5-minute measurement.
    Type of Medium: Online Resource
    ISSN: 2041-1723
    URL: Article
    DOI: 10.1038/s41467-021-24001-2
    Language: English
    Publisher: Springer Science and Business Media LLC
    Publication Date: 2021
    detail.hit.zdb_id: 2553671-0
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  • 6
    Online Resource
    Online Resource
    Middle East Respiratory Syndrome Coronavirus Spike Protein Is Not Activated Directly by Cellular Furin during Viral Entry into Target Cells
    American Society for Microbiology ; 2018
    In:  Journal of Virology Vol. 92, No. 19 ( 2018-10)
    Details
    In: Journal of Virology, American Society for Microbiology, Vol. 92, No. 19 ( 2018-10)
    Abstract: Middle East respiratory syndrome coronavirus (MERS-CoV) utilizes host cellular proteases to enter cells. A previous report shows that furin, which is distributed mainly in the Golgi apparatus and cycled to the cell surface and endosomes, proteolytically activates the MERS-CoV spike (S) protein following receptor binding to mediate fusion between the viral and cellular membranes. In this study, we reexamined furin usage by MERS-CoV using a real-time PCR-based virus cell entry assay after inhibition of cellular proteases. We found that the furin inhibitor dec-RVKR-CMK blocked entry of MERS-CoV harboring an S protein lacking furin cleavage sites; it even blocked entry into furin-deficient LoVo cells. In addition, dec-RVKR-CMK inhibited not only the enzymatic activity of furin but also those of cathepsin L, cathepsin B, trypsin, papain, and TMPRSS2. Furthermore, a virus cell entry assay and a cell-cell fusion assay provided no evidence that the S protein was activated by exogenous furin. Therefore, we conclude that furin does not play a role in entry of MERS-CoV into cells and that the inhibitory effect of dec-RVKR-CMK is specific for TMPRSS2 and cathepsin L rather than furin. IMPORTANCE Previous studies using the furin inhibitor dec-RVKR-CMK suggest that MERS-CoV utilizes a cellular protease, furin, to activate viral glycoproteins during cell entry. However, we found that dec-RVKR-CMK inhibits not only furin but also other proteases. Furthermore, we found no evidence that MERS-CoV uses furin. These findings suggest that previous studies in the virology field based on dec-RVKR-CMK should be reexamined carefully. Here we describe appropriate experiments that can be used to assess the effect of protease inhibitors on virus cell entry.
    Type of Medium: Online Resource
    ISSN: 0022-538X , 1098-5514
    URL: Article
    DOI: 10.1128/JVI.00683-18
    Language: English
    Publisher: American Society for Microbiology
    Publication Date: 2018
    detail.hit.zdb_id: 1495529-5
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  • 7
    Online Resource
    Online Resource
    S1 Subunit of Spike Protein from a Current Highly Virulent Porcine Epidemic Diarrhea Virus Is an Important Determinant of Virulence in Piglets
    MDPI AG ; 2018
    In:  Viruses Vol. 10, No. 9 ( 2018-08-30), p. 467-
    Details
    In: Viruses, MDPI AG, Vol. 10, No. 9 ( 2018-08-30), p. 467-
    Abstract: Base on the sequence of S genes, which encode spike proteins, we previously identified three different types (North American, S INDEL, and S large-DEL types) of porcine epidemic diarrhea virus (PEDV) that have re-emerged in Japan since 2013. Based on experimental infections with the North American and S large-DEL types, we also hypothesized that PEDV virulence may be linked to the S1 subunit of the S protein. To test this hypothesis, we have now assayed in gnotobiotic piglets various recombinant PEDVs generated by reverse genetics. Piglets inoculated with CV777 maintained in National Institute of Animal Health, along with piglets infected with a recombinant form of the same virus, developed subclinical to mild diarrhea. In contrast, severe watery diarrhea, dehydration, weight loss, astasia, and high mortality were observed in piglets inoculated with recombinant strains in which the S gene was partially or fully replaced with corresponding sequences from the highly virulent Japanese PEDV isolate OKN-1/JPN/2013. Indeed, symptoms resembled those in piglets inoculated with the OKN-1/JPN/2013, and were especially pronounced in younger piglets. Collectively, the data demonstrate that the S1 subunit of the S protein is an important determinant of PEDV virulence, and advance development of new vaccine candidate.
    Type of Medium: Online Resource
    ISSN: 1999-4915
    URL: Article
    DOI: 10.3390/v10090467
    Language: English
    Publisher: MDPI AG
    Publication Date: 2018
    detail.hit.zdb_id: 2516098-9
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  • 8
    Online Resource
    Online Resource
    Neutralizing and binding activities against SARS‐CoV ‐1/2, MERS‐CoV , and human coronaviruses 229E and OC43 by normal human intravenous immunoglobulin derived from healthy donors in Japan
    Wiley ; 2021
    In:  Transfusion Vol. 61, No. 2 ( 2021-02), p. 356-360
    Details
    In: Transfusion, Wiley, Vol. 61, No. 2 ( 2021-02), p. 356-360
    Abstract: There are several types of coronaviruses that infect humans and cause disease. The latest is severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2), which is an emerging global threat with no current effective treatment. Normal intravenous immunoglobulin (N‐IVIG) has been administered to coronavirus disease 2019 (COVID‐19) patients to control severe inflammation and the cellular immune response. However, the neutralizing activity of N‐IVIG against SARS‐CoV‐2 has not yet been fully evaluated. The aim of this study was to determine whether N‐IVIG manufactured before the start of the COVID‐19 pandemic contained IgG antibodies against the circulating human coronaviruses (HCoVs) that cross‐react with the highly pathogenic coronaviruses SARS‐CoV‐1, Middle East respiratory syndrome coronavirus (MERS‐CoV), and SARS‐CoV‐2. No cases of SARS‐CoV‐1 or MERS‐CoV have been reported in Japan. Study Design and Methods The neutralizing and binding activities of N‐IVIG against SARS‐CoV‐1, MERS‐CoV, SARS‐CoV‐2, HCoV 229E, and HCoV OC43 were evaluated. Nine N‐IVIG lots manufactured between 2000 and 2018, derived from donors in Japan, were tested. Binding activity was evaluated by indirect immunofluorescence assay. Results None of the N‐IVIG lots tested displayed neutralizing or binding activity against SARS‐CoV‐1, MERS‐CoV, or SARS‐CoV‐2. However, they displayed substantial neutralizing and binding activity against HCoV OC43 and weak neutralizing and substantial binding activity against HCoV 229E. Conclusion N‐IVIG derived from healthy donors in Japan before the start of the COVID‐19 pandemic had no direct effect against SARS‐CoV‐2. Further studies are warranted to determine the effects of N‐IVIG manufactured after the start of the COVID‐19 pandemic against SARS‐CoV‐2.
    Type of Medium: Online Resource
    ISSN: 0041-1132 , 1537-2995
    URL: Issue
    URL: Article
    DOI: 10.1111/trf.v61.2
    DOI: 10.1111/trf.16161
    Language: English
    Publisher: Wiley
    Publication Date: 2021
    detail.hit.zdb_id: 2018415-3
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