Extracellular vesicles containing ACE2 efficiently prevent infection by SARS‐CoV‐2 Spike protein‐containing virus
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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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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 Statement not detected. Randomization not detected. Blinding not detected. Power Analysis not detected. Sex as a biological variable not detected. Cell Line Authentication not detected. Table 2: Resources
Antibodies Sentences Resources Antibodies for WB were anti-human: ACE2 (clone EPR4435, Abcam 108252), anti-human: ACE2suggested: NoneSecondary antibodies included HRP-conjugated goat anti-rabbit IgG (H+L) (Jakson 111-035-144) anti-rabbit IgGsuggested: NoneExperimental Models: Cell Lines Sentences Resources 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-2suggested: NoneCalu-3suggested: ATCC Cat# …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 Statement not detected. Randomization not detected. Blinding not detected. Power Analysis not detected. Sex as a biological variable not detected. Cell Line Authentication not detected. Table 2: Resources
Antibodies Sentences Resources Antibodies for WB were anti-human: ACE2 (clone EPR4435, Abcam 108252), anti-human: ACE2suggested: NoneSecondary antibodies included HRP-conjugated goat anti-rabbit IgG (H+L) (Jakson 111-035-144) anti-rabbit IgGsuggested: NoneExperimental Models: Cell Lines Sentences Resources 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-2suggested: NoneCalu-3suggested: 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). Calu3suggested: None293FT 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. 293FTsuggested: 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-TMPRSS2suggested: NoneInfectivity 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. Caco2suggested: NoneSoftware and Algorithms Sentences Resources Data was analyzed using FlowJo software. FlowJosuggested: (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.
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