Platelet-RBC interaction mediated by FasL/FasR induces procoagulant activity important for thrombosis
Christoph Klatt, … , Malte Kelm, Margitta Elvers
Christoph Klatt, … , Malte Kelm, Margitta Elvers
Published June 28, 2018
Citation Information: J Clin Invest. 2018;128(9):3906-3925. https://doi.org/10.1172/JCI92077.
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Research Article Cell biology Vascular biology

Platelet-RBC interaction mediated by FasL/FasR induces procoagulant activity important for thrombosis

Abstract

Red blood cells (RBCs) influence rheology, and release ADP, ATP, and nitric oxide, suggesting a role for RBCs in hemostasis and thrombosis. Here, we provide evidence for a significant contribution of RBCs to thrombus formation. Anemic mice showed enhanced occlusion times upon injury of the carotid artery. A small population of RBCs was located to platelet thrombi and enhanced platelet activation by a direct cell contact via the FasL/FasR (CD95) pathway known to induce apoptosis. Activation of platelets in the presence of RBCs led to platelet FasL exposure that activated FasR on RBCs responsible for externalization of phosphatidylserine (PS) on the RBC membrane. Inhibition or genetic deletion of either FasL or FasR resulted in reduced PS exposure of RBCs and platelets, decreased thrombin generation, and reduced thrombus formation in vitro and protection against arterial thrombosis in vivo. Direct cell contacts between platelets and RBCs via FasL/FasR were shown after ligation of the inferior vena cava (IVC) and in surgical specimens of patients after thrombectomy. In a flow restriction model of the IVC, reduced thrombus formation was observed in FasL–/– mice. Taken together, our data reveal a significant contribution of RBCs to thrombosis by the FasL/FasR pathway.

Authors

Christoph Klatt, Irena Krüger, Saskia Zey, Kim-Jürgen Krott, Martina Spelleken, Nina Sarah Gowert, Alexander Oberhuber, Lena Pfaff, Wiebke Lückstädt, Kerstin Jurk, Martin Schaller, Hadi Al-Hasani, Jürgen Schrader, Steffen Massberg, Konstantin Stark, Hubert Schelzig, Malte Kelm, Margitta Elvers

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Figure 1

RBCs are essential for 3-dimensional thrombus formation under flow ex vivo.

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RBCs are essential for 3-dimensional thrombus formation under flow ex vi...
Human whole blood or PRP supplemented with different concentrations of RBCs was perfused over a collagen-coated surface at indicated shear rates. The effects of blood components on thrombus formation, as indicated by mean surface coverage, were determined. Data are expressed as arithmetic means. (A) Representative phase-contrast images at the end of the perfusion period. Scale bar: 100 μm. (B) Quantification of surface coverage with increasing RBC concentrations (from 1 × 106 to 4 × 106 RBCs/μl, left to right) added to PRP in comparison with whole blood. Data are given for 3 shear rates of 150 s–1 (black column), 1,000 s–1 (gray column), and 1,700 s–1 (white column). Blood components as indicated below and shear rates as shown on the left. (C) Effect of WBCs on thrombus formation under flow. Representative images of surface coverage of whole blood and PRP in the absence and presence of WBCs (7 × 103 WBCs/μl and >15 × 104 WBCs/μl, respectively). Scale bar: 50 μm. (D) Quantitative analysis of surface coverage per visual field. (E and F) Thrombus formation using PRP and fixed RBCs. Bar graphs depict mean values ± SEM. n = 3, all experiments. ***P < 0.001 by Student’s t test. PRP, platelet-rich plasma; RBC, red blood cell.