Alpha-1 antitrypsin inhibits SARS-CoV-2 infection

  1. Lukas Wettstein
  2. Carina Conzelmann
  3. Janis A. Müller
  4. Tatjana Weil
  5. Rüdiger Groß
  6. Maximilian Hirschenberger
  7. Alina Seidel
  8. Susanne Klute
  9. Fabian Zech
  10. Caterina Prelli Bozzo
  11. Nico Preising
  12. Giorgio Fois
  13. Robin Lochbaum
  14. Philip Knaff
  15. Volker Mailänder
  16. Ludger Ständker
  17. Dietmar Rudolf Thal
  18. Christian Schumann
  19. Steffen Stenger
  20. Alexander Kleger
  21. Günter Lochnit
  22. Konstantin Sparrer
  23. Frank Kirchhoff
  24. Manfred Frick
  25. Jan Münch

Reviewed by

Read the full article

Listed in

Log in to save this article

Abstract

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) causes coronavirus disease 2019 (COVID-19). To identify factors of the respiratory tract that suppress SARS-CoV-2, we screened a peptide/protein library derived from bronchoalveolar lavage, and identified α1-antitrypsin (α1-AT) as specific inhibitor of SARS-CoV-2. α1-AT targets the viral spike protein and blocks SARS-CoV-2 infection of human airway epithelium at physiological concentrations. Our findings show that endogenous α1-AT restricts SARS-CoV-2 and repurposes α1-AT-based drugs for COVID-19 therapy.

Article activity feed

  1. ScreenIT

    SciScore for 10.1101/2020.07.02.183764: (What is this?)

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

    Table 1: Rigor

    Institutional Review Board StatementIRB: Ethic statements: Ethical approval for the generation of peptide libraries from lungs and BAL was obtained from the Ethics Committee of Ulm University (application numbers 274/12 and 324/12).
    Randomizationnot detected.
    Blindingnot detected.
    Power Analysisnot detected.
    Sex as a biological variablenot detected.
    Cell Line Authenticationnot detected.

    Table 2: Resources

    Antibodies
    SentencesResources
    Protein gel was either stained with Coomassie G-250 (GelCode™ Blue Stain) or applied to western blotting of α1-AT with a polyclonal anti-α1-AT antibody (Proteintech 16382-1-AP).
    anti-α1-AT
    suggested: None
    The primary antibody was detected with labeled anti-rabbit secondary antibody and imaged with Odysey Infrared Imager (Licor)
    anti-rabbit
    suggested: None
    Subsequently cells were washed twice with PBS and incubated for 1 h at room temperature in PBS, 0.2% saponin and 10% FCS containing AlexaFluor 488-labelled anti-rabbit, AlexaFluor 468-labelled anti-mouse and AlexaFluor 647-labelled anti-rat secondary antibody, respectively (all 1:500; Thermo Scientific) and DAPI + phalloidin AF 405 (1: 5,000; Thermo Scientific).
    anti-mouse
    suggested: (R and D Systems Cat# AF-405-NA, RRID:AB_354477)
    anti-rat
    suggested: None
    Experimental Models: Cell Lines
    SentencesResources
    Cell culture: Unless stated otherwise, HEK293T cells were cultivated in DMEM supplemented with 10 % fetal calf serum (FCS), 2 mM L-glutamine, 100 U/ml penicillin and 100 mg/ml streptomycin.
    HEK293T
    suggested: None
    Cell viability assay: To assess cytotoxicity of α1-AT and camostat mesylate, 10,000 Caco2 cells were seeded in 100 μl medium in a 96-well flat bottom plate.
    Caco2
    suggested: None
    To this end, 20,000 TMPRSS2-expressing Vero E6 cells were seeded in 96 well plates in 100 μl respective medium.
    Vero E6
    suggested: None
    Software and Algorithms
    SentencesResources
    For data processing and instrument control the Compass 1.4 software package consisting of FlexControl 4.4, FlexAnalysis 3.4 4, Sequence Editor and BioTools 3.2 and ProteinScape 3.1.
    BioTools
    suggested: (MSU Mass Spectrometry Facility, RRID:SCR_012482)
    ProteinScape
    suggested: None
    Database search: Proteins were identified by MASCOT peptide mass fingerprint search (http://www.matrixscience.com) using the Uniprot Human database (version 20200226, 210438 sequence entries; p<0.05).
    http://www.matrixscience.com
    suggested: (MatrixScience, RRID:SCR_000307)
    Non-linear regression and statistics: Analysis was performed using GraphPad Prism version 8.4.2. Calculation of IC50 values via non-linear regression was performed using normalized response-variable slope equation.
    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 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

    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 10.1101/2020.07.02.183764v1 on bioRxiv