Antiviral roles of interferon regulatory factor (IRF)-1, 3 and 7 against human coronavirus infection

  1. Danyang Xu
  2. Joseph K. Sampson Duncan
  3. Maria Licursi
  4. Jin Gohda
  5. Yasushi Kawaguchi
  6. Kensuke Hirasawa

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Abstract

Interferon regulatory factors (IRFs) are key elements of antiviral innate responses that regulate transcription of interferons (IFNs) and IFN-stimulated genes (ISGs). As many human coronaviruses are known to be sensitive to IFN, antiviral roles of IRFs are yet to be fully understood. TypeI or II IFN treatment protected MRC5 cells from infection of human coronavirus 229E, but not human coronavirus OC43. Infection of 229E or OC43 efficiently upregulated ISGs, indicating that antiviral transcription is not suppressed during their infection. Antiviral IRFs, IRF1, IRF3 and IRF7, were activated in cells infected with 229E, OC43 or severe acute respiratory syndrome-associated coronavirus 2 (SARS-CoV-2). RNAi knockdown and overexpression of the IRFs demonstrated that IRF1 and IRF3 have antiviral property against OC43 while only IRF3 and IRF7 are effective to restrict 229E infection. Our study demonstrates that IRF3 plays critical roles against infection of human coronavirus 229E and OC43, which may be an anti-human coronavirus therapeutic target.

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  1. ScreenIT

    SciScore for 10.1101/2022.03.24.485591: (What is this?)

    Please note, not all rigor criteria are appropriate for all manuscripts.

    Table 1: Rigor

    Ethicsnot detected.
    Sex as a biological variablenot detected.
    Randomizationnot detected.
    Blindingnot detected.
    Power Analysisnot detected.
    Cell Line Authenticationnot detected.

    Table 2: Resources

    Antibodies
    SentencesResources
    Antibodies used in this study include: IRF3, phospho-IRF3, IRF7, phsopho-STAT1 (Cell signalling technology), IRF1 (BD Transduction Laboratories), GAPDH (Santa Cruze Biotechnology), 229E N protein (Ingenasa), OC43 N protein (Milipore), SARS-CoV-2 N protein (Sino Biological).
    IRF3
    suggested: None
    phospho-IRF3, IRF7
    suggested: None
    phsopho-STAT1
    suggested: None
    IRF1
    suggested: None
    GAPDH
    suggested: None
    Experimental Models: Cell Lines
    SentencesResources
    Cells, viruses, and reagents: Human lung fibroblast cell line MRC5, human lung cancer cell line H1299, human lung cancer cell line Calu-3, mouse fibroblast cell line L929, human coronaviruses HCoV-OC43 and HCoV-229E were obtained from the American Type Culture Collection (ATCC; Manassas, VA, USA).
    MRC5
    suggested: None
    H1299
    suggested: None
    Calu-3
    suggested: None
    HCoV-229E
    suggested: JCRB Cat# JCRB1838, RRID:CVCL_B3M4)
    VSV was amplified and titrated by plaque assay using L929 cells as described previously (47).
    L929
    suggested: ECACC Cat# 86032004, RRID:CVCL_4238)
    The SARS-CoV-2 isolate (UT-NCGM02/Human/2020/Tokyo) (49) was propagated in VeroE6-TMPRSS2 cells in DMEM containing 5% heat-inactivated FBS at 37 °C in 5 % CO2.
    VeroE6-TMPRSS2
    suggested: JCRB Cat# JCRB1819, RRID:CVCL_YQ49)
    Recombinant DNA
    SentencesResources
    pcDNA3-IRF3 was purchased from Addgene.
    pcDNA3-IRF3
    suggested: None
    For overexpression of IRFs, cells in 24-well plates (4×104 cells/well) were transfected with control pcDNA3, pcDNA-IRF1, pcDNA-IRF3 or pcDNA-IRF7 using Lipofectamine 3000 Transfection Reagent (Life Technologies) and 24 hours later challenged with or without 229E or OC43.
    pcDNA3
    suggested: RRID:Addgene_15475)
    pcDNA-IRF1
    suggested: None
    pcDNA-IRF3
    suggested: None
    pcDNA-IRF7
    suggested: None
    Software and Algorithms
    SentencesResources
    Statistical analysis: Two-way ANOVA followed by Sidak’s post hoc test was performed using GraphPad Prism 6.0 software.
    GraphPad Prism
    suggested: (GraphPad Prism, RRID:SCR_002798)

    Results from OddPub: We did not detect open data. We also did not detect open code. Researchers are encouraged to share open data when possible (see Nature blog).

    Results from LimitationRecognizer: An explicit section about the limitations of the techniques employed in this study was not found. We encourage authors to address study limitations.

    Results from TrialIdentifier: No clinical trial numbers were referenced.

    Results from Barzooka: We did not find any issues relating to the usage of bar graphs.

    Results from JetFighter: We did not find any issues relating to colormaps.

    Results from rtransparent:
    • Thank you for including a conflict of interest statement. Authors are encouraged to include this statement when submitting to a journal.
    • Thank you for including a funding statement. Authors are encouraged to include this statement when submitting to a journal.
    • No protocol registration statement was detected.

    Results from scite Reference Check: We found no unreliable references.

    About SciScore

    SciScore is an automated tool that is designed to assist expert reviewers by finding and presenting formulaic information scattered throughout a paper in a standard, easy to digest format. SciScore checks for the presence and correctness of RRIDs (research resource identifiers), and for rigor criteria such as sex and investigator blinding. For details on the theoretical underpinning of rigor criteria and the tools shown here, including references cited, please follow this link.

  2. Version published to 10.1101/2022.03.24.485591 on bioRxiv