Aberrant glycosylation of anti-SARS-CoV-2 spike IgG is a prothrombotic stimulus for platelets
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Abstract
A subset of patients with coronavirus disease 2019 (COVID-19) become critically ill, suffering from severe respiratory problems and also increased rates of thrombosis. The causes of thrombosis in severely ill patients with COVID-19 are still emerging, but the coincidence of critical illness with the timing of the onset of adaptive immunity could implicate an excessive immune response. We hypothesized that platelets might be susceptible to activation by anti–severe acute respiratory syndrome coronavirus 2 (anti-SARS-CoV-2) antibodies and might contribute to thrombosis. We found that immune complexes containing recombinant SARS-CoV-2 spike protein and anti-spike immunoglobulin G enhanced platelet-mediated thrombosis on von Willebrand factor in vitro, but only when the glycosylation state of the Fc domain was modified to correspond with the aberrant glycosylation previously identified in patients with severe COVID-19. Furthermore, we found that activation was dependent on FcγRIIA, and we provide in vitro evidence that this pathogenic platelet activation can be counteracted by the therapeutic small molecules R406 (fostamatinib) and ibrutinib, which inhibit tyrosine kinases Syk and Btk, respectively, or by the P2Y12 antagonist cangrelor.
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SciScore for 10.1101/2021.03.26.437014: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement Consent: Blood Preparation: Blood samples were obtained from healthy donors that had given informed consent and using procedures approved by the University of Reading Research Ethics Committee and collected into vacutainers containing 3.8% (w/v) sodium
IRB: Blood Preparation: Blood samples were obtained from healthy donors that had given informed consent and using procedures approved by the University of Reading Research Ethics Committee and collected into vacutainers containing 3.8% (w/v) sodiumRandomization not detected. Blinding not detected. Power Analysis not detected. Sex as a biological variable not detected. Cell Line Authentication not detected. Table 2: Resources
… SciScore for 10.1101/2021.03.26.437014: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement Consent: Blood Preparation: Blood samples were obtained from healthy donors that had given informed consent and using procedures approved by the University of Reading Research Ethics Committee and collected into vacutainers containing 3.8% (w/v) sodium
IRB: Blood Preparation: Blood samples were obtained from healthy donors that had given informed consent and using procedures approved by the University of Reading Research Ethics Committee and collected into vacutainers containing 3.8% (w/v) sodiumRandomization 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 The slides were then washed and treated with 10μg/ml COVA1-18 recombinant, monoclonal anti-SARS-COV-2 antibodies (see above) for 1h at 37°C and then 2U/ml vWF for 1h. anti-SARS-COV-2suggested: NoneFlow cytometry measurement of fibrinogen binding: Measurements of fibrinogen binding were performed using washed platelets pretreated with immune complexes, created by incubating 5μg/ml recombinant SARS-COV-2 spike protein with 10μg/ml COVA1-18 for 60 minutes at 37°C, and then stimulated with 1 μg/mL CRP, 10μM ADP or 1μM TRAP-6 in the presence of fluorescein isothiocyanate–conjugated polyclonal rabbit anti-fibrinogen antibody (Agilent Technologies LDA UK Limited, Cheadle, United Kingdom), and then incubated for 20 minutes in the dark. anti-fibrinogensuggested: NoneExperimental Models: Cell Lines Sentences Resources Recombinant anti SARS-CoV-2 IgG: COVA1-18 IgG was produced in 293F cells as described previously [13]. 293Fsuggested: RRID:CVCL_D615)Software and Algorithms Sentences Resources Statistical methods: Statistical testing as described in figure legends and the results section were performed using GraphPad Prism Software (GraphPad, La Jolla, CA). GraphPad Prismsuggested: (GraphPad Prism, RRID:SCR_002798)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: We found the following clinical trial numbers in your paper:
Identifier Status Title NCT04579393 Active, not recruiting Fostamatinib for Hospitalized Adults With COVID-19 NCT04581954 Recruiting Inflammatory Signal Inhibitors for COVID-19 (MATIS) 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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