Abstract Number: PB0369
Meeting: ISTH 2021 Congress
Background: The Fibrin Polymerization (FP) and Coag-Lysis (CL) assays are fully automated for the turbidimetric measurement of the Fibrin Polymerization (1) and Coagulation/Fibrinolysis (2) profiles. Neutrophil Extracellular Traps (NETs) are known to modify the Fibrin structure and render it resistant to destruction (3).
Aims: To analyse the in vitro effect of NETs in FP and CL assays using spiking, inhibition and cells activation models.
Methods: DNA or Histone 3 (H3) (70µg/mL; 50µg/mL) and their inhibition with DNAse or H3 inhibitor (140U; 50µg/mL) were respectively tested in normal pool plasma on FP and CL using PPP Reagent Low® with (CL) or without (FP) tissue Plasminogen Activator. Whole blood and isolated neutrophils from healthy donors were activated with LPS or PMA and Flow Cytometry analysis of leukocytes and FP/CL determinations were conducted. The presence of DNA from NETs was evidenced by confocal microscopy using Dapi and Picogreen.
Results: DNA had pro-coagulant and anti-fibrinolytic effects in FP and CL, as opposed to H3 which had anti-coagulant, clot stabilisation and pro-fibrinolytic effects.
The effect of DNA was partially reversed by DNase, while Fibrin/Lysis parameters were completely restored with the H3 inhibitor.
In activation models, DNA was released and DNA-like effect was obtained in FP and CL.
A decreased size and DNA related fluorescence of neutrophils and an increased granularity, DNA fluorescence and TF/HLA-DR expression of monocytes, testified to their activation.
Conclusions: DNA released from cells activation modified FP and CL profiles, down regulating clot lysis. H3 effect was observed in spiking model only, as opposed to DNA’s effect on FP and CL profiles due to its positive charges as shown in inhibition model. Therapeutic strategies could be optimized on these bases to enhance fibrinolysis in clots containing DNA and histones.
(1) Contant G. RPTH 2017.
(2) Pieters M. JTH 2018.
(3) Longstaff C. J. Biol. Chem. 2013.
To cite this abstract in AMA style:Biatougou NM-, Mas F, Montpellier B, Mirshahi S, Soria J, Contant G. Effect of NETS from Different Models on Fibrin Polymerization Profile and Coagulation-fibrinolysis Balance [abstract]. Res Pract Thromb Haemost. 2021; 5 (Suppl 2). https://abstracts.isth.org/abstract/effect-of-nets-from-different-models-on-fibrin-polymerization-profile-and-coagulation-fibrinolysis-balance/. Accessed July 1, 2022.
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