LY6E Restricts Entry of Human Coronaviruses, Including Currently Pandemic SARS-CoV-2

  1. Xuesen Zhao
  2. Shuangli Zheng
  3. Danying Chen
  4. Mei Zheng
  5. Xinglin Li
  6. Guoli Li
  7. Hanxin Lin
  8. Jinhong Chang
  9. Hui Zeng
  10. Ju-Tao Guo

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Abstract

Virus entry into host cells is one of the key determinants of host range and cell tropism and is subjected to the control of host innate and adaptive immune responses. In the last decade, several interferon-inducible cellular proteins, including IFITMs, GILT, ADAP2, 25CH, and LY6E, had been identified to modulate the infectious entry of a variety of viruses. Particularly, LY6E was recently identified as a host factor that facilitates the entry of several human-pathogenic viruses, including human immunodeficiency virus, influenza A virus, and yellow fever virus. Identification of LY6E as a potent restriction factor of coronaviruses expands the biological function of LY6E and sheds new light on the immunopathogenesis of human coronavirus infection.

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  1. Version published to 10.1128/jvi.00562-20
  2. ScreenIT

    SciScore for 10.1101/2020.04.02.021469: (What is this?)

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

    Table 1: Rigor

    Institutional Review Board Statementnot detected.
    Randomizationnot detected.
    Blindingnot detected.
    Power Analysisnot detected.
    Sex as a biological variablenot detected.
    Cell Line Authenticationnot detected.

    Table 2: Resources

    Antibodies
    SentencesResources
    Antibodies: Monoclonal antibody against FLAG tag (ANTI-FLAG M2) and β-Actin were purchased from Sigma (Cat.No. F1804 and A2228, respectively).
    Antibodies
    suggested: (Sigma-Aldrich Cat# A2228, RRID:AB_476697)
    ANTI-FLAG
    suggested: (Sigma-Aldrich Cat# F1804, RRID:AB_262044)
    β-Actin
    suggested: (Sigma-Aldrich Cat# F1804, RRID:AB_262044)
    A2228
    suggested: (Sigma-Aldrich Cat# A2228, RRID:AB_476697)
    Monoclonal antibody against human IFITM1 (Cat.No. 60047-1), rabbit polyclonal antibody against human IFITM3 (Cat.No. 11714-1-AP), which also efficiently recognizes IFITM2 and weakly cross-reacts with IFITM1, were purchased from Proteintech Group, Inc.
    human IFITM1
    suggested: (Novus Cat# NB 600-471, RRID:AB_535506)
    human IFITM3
    suggested: (Proteintech Cat# 11714-1-AP, RRID:AB_2295684)
    IFITM1
    suggested: None
    Mouse monoclonal antibody against HCoV-OC43 nucleocapsid (NP) protein was purchased from Millipore (Cat.No. MAB9012).
    HCoV-OC43 nucleocapsid (NP) protein
    suggested: None
    HCoV-OC43 nucleocapsid (NP
    suggested: None
    After permeabilization with 0.1% Triton X-100, the cells were stained with a monoclonal antibody (541-8F) recognizing HCoV-OC43 NP protein.
    HCoV-OC43 NP protein.
    suggested: None
    The bound antibodies were visualized by using Alexa Fluor 488-labeled (green) goat anti-mouse IgG or Alexa Fluor 555-labeled (red) goat anti-mouse IgG, Cell nuclei were counterstained with DAPI.
    anti-mouse IgG
    suggested: None
    Experimental Models: Cell Lines
    SentencesResources
    Cell culture: Human hepatoma cell lines HepG2 and C3A, a sub-clone of HepG2 (ATCC HB-8065) were purchased from ATCC and cultured in DMEM/F12 medium supplemented with 10% heat-inactivated fetal bovine serum (FBS) (Invitrogen).
    HepG2
    suggested: ATCC Cat# HB-8065, RRID:CVCL_0027)
    Lung cancer cell line A549 were obtained from ATCC and maintained in DMEM supplemented with 10% FBS.
    A549
    suggested: None
    GP2-293 and Lenti-X 293T cell Lines were purchased from Clontech and cultured in DMEM supplemented with 10% FBS and 1 mM Sodium pyruvate (Invitrogen).
    293T
    suggested: None
    Flp-In TREx 293 cells were purchased from Invitrogen and maintained in DMEM supplemented with 10% FBS, 10 μg/ml blasticidin (Invitrogen) and 100 µg/ml Zeocin (Invivogen) (64).
    Flp-In TREx 293
    suggested: RRID:CVCL_U427)
    Flp-In TREx 293-derived cell lines expressing LY6E, GILT, ADAP2, or IFITM3 were cultured in DMEM supplemented with 10% FBS, 5 μg/ml blasticidin and 250 μg/ml hygromycin. Viruses: HCoV-OC43 (strain VR1558) were purchased from ATCC and amplified in HCT-8 cells according to the instruction from ATCC.
    HCT-8
    suggested: ICLC Cat# HTL99005, RRID:CVCL_2478)
    Establishment of cell lines stably expressing wild-type and mutant IFITM or LY6E proteins or shRNA: HepG2, C3A or A549 cells in each well of 6-well plates were incubated with 2 ml of Opti-MEM medium containing pseudotyped retroviruses and centrifuged at 20 °C for 30 minutes at 4,000×g.
    C3A
    suggested: None

    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 found bar graphs of continuous data. We recommend replacing bar graphs with more informative graphics, as many different datasets can lead to the same bar graph. The actual data may suggest different conclusions from the summary statistics. For more information, please see Weissgerber et al (2015).

    Results from JetFighter: Please consider improving the rainbow (“jet”) colormap(s) used on page 33. At least one figure is not accessible to readers with colorblindness and/or is not true to the data, i.e. not perceptually uniform.

    Results from rtransparent:
    • No conflict of interest statement was detected. If there are no conflicts, we encourage authors to explicit state so.
    • 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.

    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.

  3. Version published to 10.1101/2020.04.02.021469v1 on bioRxiv