Extracellular vesicles containing ACE2 efficiently prevent infection by SARS‐CoV‐2 Spike protein‐containing virus

  1. Federico Cocozza
  2. Nathalie Névo
  3. Ester Piovesana
  4. Xavier Lahaye
  5. Julian Buchrieser
  6. Olivier Schwartz
  7. Nicolas Manel
  8. Mercedes Tkach
  9. Clotilde Théry
  10. Lorena Martin‐Jaular

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Abstract

SARS‐CoV‐2 entry is mediated by binding of the spike protein (S) to the surface receptor ACE2 and subsequent priming by host TMPRSS2 allowing membrane fusion. Here, we produced extracellular vesicles (EVs) exposing ACE2 and demonstrate that ACE2‐EVs are efficient decoys for SARS‐CoV‐2 S protein‐containing lentivirus. Reduction of infectivity positively correlates with the level of ACE2, is much more efficient than with soluble ACE2 and further enhanced by the inclusion of TMPRSS2.

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  1. Version published to 10.1002/jev2.12050
  2. ScreenIT

    SciScore for 10.1101/2020.07.08.193672: (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 for WB were anti-human: ACE2 (clone EPR4435, Abcam 108252),
    anti-human: ACE2
    suggested: None
    Secondary antibodies included HRP-conjugated goat anti-rabbit IgG (H+L) (Jakson 111-035-144)
    anti-rabbit IgG
    suggested: None
    Experimental Models: Cell Lines
    SentencesResources
    Cells: Human Caco-2 (HTB-37) and Calu-3 (HTB-55) were purchased from ATCC and maintained at 37°C in a humidified atmosphere with 5% CO2.
    Caco-2
    suggested: None
    Calu-3
    suggested: ATCC Cat# HTB-55, RRID:CVCL_0609)
    Caco2 and Calu3 cells were cultured in DMEM (Sigma) supplemented with 10% FBS (Gibco), 100U/ml penicillin-streptomycin (Thermo Fisher Scientific) and non-essential aminoacids (Thermo Fisher Scientific).
    Calu3
    suggested: None
    293FT cells were cultured in DMEM medium (Sigma) supplemented with 10% FBS (Eurobio) and 100U/ml penicillin-streptomycin (Thermo Fisher Scientific). 293FT-mock, 293FT-ACE2 and 293FT-ACE2-TMPRSS2 cells were generated by stable double transduction with pTRIP-SFFV-tagBFP-2A and pTRIP-SFFV-TagRFP657-2A, pTRIP-SFFV-tagBFP-2A-hACE2 and pTRIP-SFFV-TagRFP657-2A, or pTRIP-SFFV-tagBFP-2A-hACE2 and pTRIP-SFFV-TagRFP657-2A-TMPRSS2, respectively.
    293FT
    suggested: ATCC Cat# PTA-5077, RRID:CVCL_6911)
    EV isolation by Size-Exclusion Chromatography (SEC): 239FT-mock, 293FT-ACE2 and 293FT-ACE2-TMPRSS2 cells were cultured in serum EV-depleted medium for 24h.
    293FT-ACE2-TMPRSS2
    suggested: None
    Infectivity Assay: 10,000-20,000 293FT-ACE2, Caco2 and Calu3 cells were seeded in a 96 well plate and after 6 h infected with SARS-CoV-2 S-pseudotyped virus in EV-depleted medium.
    Caco2
    suggested: None
    Software and Algorithms
    SentencesResources
    Data was analyzed using FlowJo software.
    FlowJo
    suggested: (FlowJo, RRID:SCR_008520)

    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.

    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 10.1101/2020.07.08.193672v1 on bioRxiv