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 6

Cell-cell contact between platelets and RBCs via FasR (CD95).

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Cell-cell contact between platelets and RBCs via FasR (CD95). (A) Effect...
(A) Effects of FasR-blocking antibody on PS exposure of RBCs in the presence of ADP-stimulated platelets. (n = 9). (B) Platelet PS exposure was determined by flow cytometry (n = 4–6). (C and D) Thrombin-induced thrombin generation in PRP supplemented with RBCs in the presence and absence of FasR-blocking antibody was measured by calibrated automated thrombogram. (C) Representative thrombin curves and (D) endogenous thrombin potential (ETP) (nM × min) are shown. (E–H) Flow chamber experiments with FasR-blocking antibody. (E) Representative images of thrombus formation. Scale bar: 50 μm. (F) Mean surface coverage per visual field (n = 5). (G and H) Effects of FasR-blocking antibody on PS exposure of RBCs (G) and platelets (H) (n = 7–8). (I and J) FeCl3-induced injury of mesenteric arterioles. WT mice were injected with FasR-blocking antibody (anti-FasR, 20 μg/mouse) and IgG (20 μg/mouse) as control. Thrombus formation was monitored by intravital microscopy. (I) Time to irreversible occlusion (right panel). (J) Representative images at indicated time points (n = 4–6/group). (K) Tail bleeding times were determined in mice (n = 5–8/group). FasR antibody was used at 10 μg/ml. Agonist concentrations: ADP (10 μM) and CRP (5 μg/ml). Data are the mean ± SEM. *P < 0.05; **P < 0.01; ***P < 0.001 by 1-way ANOVA with Tukey’s multiple-comparisons test (A and B) or Student’s t test (D–K). NS, not significant; Plts, platelets.