Upregulation of cAMP prevents antibody-mediated thrombus formation in COVID-19

Article activity feed

1. Version published to 10.1182/bloodadvances.2021005210

   Jan 10, 2022
2. 

   ScreenIT

   Dec 17, 2020

   SciScore for 10.1101/2020.12.15.20247775: (What is this?)

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

   Table 1: Rigor

   Institutional Review Board Statement	IRB: : Studies involving human material were approved by the Ethics Committee of the Medical Faculty, Eberhard-Karls University of Tuebingen, Germany, and were conducted in accordance with the declaration of Helsinki.
   Randomization	Immunofluorescence and bright field images were taken from 3-5 randomly chosen microscopic fields (x20, Olympus IX73, Olympus GmbH, Hamburg, Germany)
   Blinding	not detected.
   Power Analysis	not detected.
   Sex as a biological variable	not detected.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   Detection of phosphatidylserine exposure: To assess externalization of phosphatidylserine (PS) on the PLT surface after antibody treatment, 10 µL of PLT …

   SciScore for 10.1101/2020.12.15.20247775: (What is this?)

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

   Table 1: Rigor

   Institutional Review Board Statement	IRB: : Studies involving human material were approved by the Ethics Committee of the Medical Faculty, Eberhard-Karls University of Tuebingen, Germany, and were conducted in accordance with the declaration of Helsinki.
   Randomization	Immunofluorescence and bright field images were taken from 3-5 randomly chosen microscopic fields (x20, Olympus IX73, Olympus GmbH, Hamburg, Germany)
   Blinding	not detected.
   Power Analysis	not detected.
   Sex as a biological variable	not detected.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   Detection of phosphatidylserine exposure: To assess externalization of phosphatidylserine (PS) on the PLT surface after antibody treatment, 10 µL of PLT suspension were transferred into 100 µL of Hank’s balanced salt solution (HBSS), (137 mM NaCl, 1.25 mM CaCl2, 5.5 mM glucose, [Carl-Roth, Karlsruhe, Germany]) and incubated with 1 µL Annexin V-FITC (Immunotools, Friesoythe, Germany) for 30 min at RT in the dark.	Annexin V-FITC suggested: (Thermo Fisher Scientific Cat# BMS500FI/100, RRID:AB_2575598)
   Membranes were then incubated with primary anti-human cleaved-caspase 3 antibody (1:1000, Abcam, Cambridge, UK) and anti-human alpha-tubulin (1:1000, Cell Signaling Technology, Danvers, USA) at 4°C overnight.	anti-human cleaved-caspase 3 suggested: None anti-human alpha-tubulin suggested: None
   After washing with TBS-T buffer, the membranes were incubated with the appropriate secondary anti-rabbit or anti-mouse antibody conjugated with IRDye®680 / IRDye®800 (1:3000, LI-COR®, Lincoln, USA) for 1 h at RT.	anti-rabbit suggested: None anti-mouse suggested: None
   Assessment of the mechanisms of antibody-mediated effects on PLTs: 75 µL wPLTs were pretreated with the FcγRIIA blocking monoclonal antibody (moAb) anti-CD32 (5 µL moAb IV.3;	anti-CD32 suggested: None
   Software and Algorithms
   Sentences	Resources
   Western blots were analyzed by ImageJ software (NIH, Bethesda,	ImageJ suggested: (ImageJ, RRID:SCR_003070)
   Statistical analysis: Statistical analyses were performed using GraphPad Prism 7 (La Jolla, USA).	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: We detected the following sentences addressing limitations in the study:

   Our study is subjected to some limitations. First, as an observational, monocentric study, we cannot conclude that the reported associations between IgG antibodies and changes in activation/apoptosis markers in COVID-19 are causal for the thrombosis or specific for the disease. Second, we cannot exclude the possibility of remaining residual confounding or unmeasured potential confounders in our mechanistic studies. Third, the low number of patients does not enable a final and robust statistical analysis to assess clinical outcomes in patients with increased procoagulant PLTs. Nevertheless, data presented in this study may provide a background for future studies to dissect mechanisms related to PLT activation that are involved in the progression of COVID-19. Taken together, our study shows that IgG antibodies from patients with severe COVID-19 are able to stimulate FcγRIIA leading to the induction of procoagulant PLTs with an increased ability of clot formation. These processes are dependent on calcium and can be efficiently inhibited by cAMP inducers suggesting that ADC might represent a potentially promising target to prevent thromboembolic complications in COVID-19 disease.

   ---

   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.

   ---

   Read the original source
3. 

   Version published to 10.1101/2020.12.15.20247775v1 on medRxiv

   Dec 17, 2020