PSGL-1 inhibits the virion incorporation of SARS-CoV and SARS-CoV-2 spike glycoproteins and impairs virus attachment and infectivity

  1. Sijia He
  2. Abdul A. Waheed
  3. Brian Hetrick
  4. Deemah Dabbagh
  5. Ivan V. Akhrymuk
  6. Kylene Kehn-Hall
  7. Eric O. Freed
  8. Yuntao Wu

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Abstract

P-selectin glycoprotein ligand-1 (PSGL-1) is a cell surface glycoprotein that binds to P-, E-, and L-selectins to mediate the tethering and rolling of immune cells on the surface of the endothelium for cell migration into inflamed tissues. PSGL-1 has been identified as an interferon-γ (INF-γ)-regulated factor that restricts HIV-1 infectivity, and has recently been found to possess broad-spectrum antiviral activities. Here we report that the expression of PSGL-1 in virus-producing cells impairs the incorporation of SARS-CoV and SARS-CoV-2 spike (S) glycoproteins into pseudovirions and blocks virus attachment and infection of target cells. These findings suggest that PSGL-1 may potentially inhibit coronavirus replication in PSGL-1 + cells.

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  1. Version published to 10.1101/2020.05.01.073387v2 on bioRxiv
  2. ScreenIT

    SciScore for 10.1101/2020.05.01.073387: (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
    Virions were purified and analyzed by SDS-PAGE and western blot using antibodies against SARS-CoV spike proteins (Genetex), PSGL-1 (KPL-1 clone), or HIV-Ig to detect CA protein p24.
    PSGL-1
    suggested: None
    Cell lysates were analyzed by SDS-PAGE and western blotting with a mouse anti-HIV-1 p24 monoclonal antibody (NIH AIDS Reagent Program, 183-H12-5C) (1:1000 dilution) or anti-GAPDH goat polyclonal antibody (Abcam) (1:1000 dilution) at 4°C overnight, then washed and incubated with anti-mouse IgG, HRP-linked antibody (Cell Signaling) (1:2000 dilution) or anti-goat horseradish peroxidase-conjugated antibody (KPL) (1:2500 dilution) at room temperature for 30-60 minutes.
    anti-HIV-1
    suggested: (NIH AIDS Reagent Program Cat# 3537, RRID:AB_2832923)
    183-H12-5C
    suggested: (Dr. Bruce Chesebro and Dr. Hardy Chen - National Institute of Allergy and Infectious Diseases Cat# 183-H12-5C, RRID:AB_2819250)
    anti-GAPDH
    suggested: None
    anti-mouse IgG
    suggested: (Cayman Chemical Cat# 400002-2500, RRID:AB_328092)
    HRP-linked antibody (Cell Signaling)
    suggested: None
    anti-goat horseradish peroxidase-conjugated antibody (KPL)
    suggested: None
    Experimental Models: Cell Lines
    SentencesResources
    Both of SARS-CoV-S and SARS-CoV-2-S pseudotyped viral particles were produced in HEK293T cells.
    HEK293T
    suggested: NCBI_Iran Cat# C498, RRID:CVCL_0063)
    Viral infectivity assay: Virus particles produced in the presence of PSGL-1 or the empty vector were used to infect Vero E6 or Calu-3 cells (ATCC).
    Vero E6
    suggested: RRID:CVCL_XD71)
    Calu-3
    suggested: KCLB Cat# 30055, RRID:CVCL_0609)

    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: 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.

    About SciScore

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  3. Version published to 10.1101/2020.05.01.073387v1 on bioRxiv