IgA MAb blocks SARS-CoV-2 Spike-ACE2 interaction providing mucosal immunity
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Abstract
COVID-19 caused by SARS-CoV-2 has become a global pandemic requiring the development of interventions for the prevention or treatment to curtail mortality and morbidity. No vaccine to boost mucosal immunity or as a therapeutic has yet been developed to SARS-CoV-2. In this study we discover and characterize a cross-reactive human IgA monoclonal antibody, MAb362. MAb362 binds to both SARS-CoV and SARS-CoV-2 spike proteins and competitively blocks hACE2 receptor binding, by completely overlapping the hACE2 structural binding epitope. Furthermore, MAb362 IgA neutralizes both pseudotyped SARS-CoV and SARS-CoV-2 in human epithelial cells expressing hACE2. SARS-CoV-2 specific IgA antibodies, such as MAb362, may provide effective immunity against SARS-CoV-2 by inducing mucosal immunity within the respiratory system, a potentially critical feature of an effective vaccine.
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SciScore for 10.1101/2020.05.15.096719: (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 Isotype switching was conducted using primers designed to amplify the variable heavy chain of the IgG antibody. IgGsuggested: NoneIgG and IgA1 antibodies were transfected in Expi293 cells and purified as previously described32. IgA1suggested: NoneAfter incubation, the cell pellets were washed and then resuspened in PBS with 2% FBS and incubated with 10 µg/mL of anti-Myc antibody for 1 hour at 4°C. anti-Mycsuggested: NoneExperimental Models: Cell Lines Sentence… SciScore for 10.1101/2020.05.15.096719: (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 Isotype switching was conducted using primers designed to amplify the variable heavy chain of the IgG antibody. IgGsuggested: NoneIgG and IgA1 antibodies were transfected in Expi293 cells and purified as previously described32. IgA1suggested: NoneAfter incubation, the cell pellets were washed and then resuspened in PBS with 2% FBS and incubated with 10 µg/mL of anti-Myc antibody for 1 hour at 4°C. anti-Mycsuggested: NoneExperimental Models: Cell Lines Sentences Resources IgG and IgA1 antibodies were transfected in Expi293 cells and purified as previously described32. Expi293suggested: RRID:CVCL_D615)Flow cytometry-based receptor binding inhibition assay: Vero E6 cells were harvested with PBS containing 5 mM EDTA and aliquoted to 1×106 cells per reaction. Vero E6suggested: NoneBefore mixing with the cells, Myc-tagged SARS-CoV S1-590 or SARS-CoV-2 S1-604 was incubated with the MAb at varying concentrations for 1 hour at room temperature, then the S protein was added to the Vero cells to a final concentration of 10 nM. Verosuggested: NoneSARS-S and SARS2-S spike protein was provided in trans by co-transfection of 293T cells with pcDNA-G with pNL4-3. 293Tsuggested: NoneSoftware and Algorithms Sentences Resources Cells were washed twice then subjected to flow cytometric analysis using a MACSquant Flow Cytometer (Miltenyi Biotec) and analyzed by FlowJo (version 10). FlowJosuggested: (FlowJo, RRID:SCR_008520)The molecular modeling of MAb362 was performed through the program Modeller 9.15 using the basic modeling and forming the initial MAb362:SARS-CoV RBD complex. Modellersuggested: (MODELLER, RRID:SCR_008395)Hydrogen bonds were determined for pairs of eligible donor/acceptor atoms using criteria set by Schrodinger (Schrodinger, LLC, The PyMOL Molecular Graphics System, Version 1.3r1. 2010.). PyMOLsuggested: (PyMOL, RRID:SCR_000305)Each assay was performed in triplicate. c Statistical Analysis: Statistical calculations were performed using Prism version 7.03 (GraphPad Software, La Jolla, CA). Prismsuggested: (PRISM, RRID:SCR_005375)GraphPadsuggested: (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 did not find any issues relating to the usage of bar graphs.
Results from JetFighter: Please consider improving the rainbow (“jet”) colormap(s) used on page 14. At least one figure is not accessible to readers with colorblindness and/or is not true to the data, i.e. not perceptually uniform.
Results from rtransparent:- Thank you for including a conflict of interest statement. Authors are encouraged to include this statement when submitting to a journal.
- No funding statement was detected.
- No protocol registration statement was detected.
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