Alpha-1 antitrypsin inhibits SARS-CoV-2 infection
This article has been Reviewed by the following groups
Listed in
- Evaluated articles (ScreenIT)
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
-
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 Statement IRB: 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). 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 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-ATsuggested: NoneThe primary antibody was detected with labeled anti-rabbit secondary … 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 Statement IRB: 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). 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 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-ATsuggested: NoneThe primary antibody was detected with labeled anti-rabbit secondary antibody and imaged with Odysey Infrared Imager (Licor) anti-rabbitsuggested: NoneSubsequently 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-mousesuggested: (R and D Systems Cat# AF-405-NA, RRID:AB_354477)anti-ratsuggested: NoneExperimental Models: Cell Lines Sentences Resources 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. HEK293Tsuggested: NoneCell 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. Caco2suggested: NoneTo this end, 20,000 TMPRSS2-expressing Vero E6 cells were seeded in 96 well plates in 100 μl respective medium. Vero E6suggested: NoneSoftware and Algorithms Sentences Resources 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. BioToolssuggested: (MSU Mass Spectrometry Facility, RRID:SCR_012482)ProteinScapesuggested: NoneDatabase 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.comsuggested: (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 Prismsuggested: (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.
-