Anti-β2 Glycoprotein I Antibodies Prevent the De-activation of Platelets and Sustain their Phagocytic Clearance

Abstract

Exposure to phosphatidylserine (PS) tags dying and senescent cells for removal and identifies activated platelets. In this study we followed the fate of PS-exposing platelets in the presence of antibodies purified from Systemic Lupus Erythematosus (SLE) and primary Anti-phospholipid Syndrome (APS) patients' sera by β2GPI affinity chromatography. Thrombin-activated platelets exposed PS and associated to β2GPI. Both events were required for recognition by antibodies. Human monocyte-derived macrophages phagocytosed activated platelets only. Each macrophage internalized an average of 3.16±0.2 platelets after 60min at 37°C. Phagocytosis did not increase after longer incubations (4.65±0.26 platelets internalized by each macrophage after 300min). Recognition of platelets by anti-β2GPI antibodies significantly increased phagocytosis (P< 0.01). Upon withdrawal of thrombin, platelets downregulated PS (PS exposure t1/2: 242min) and the ability to be recognized by macrophages. Purified β2GPI bound to PS-exposing platelets (association t1/2: 250min). Phosphatidyl serine exposure and β2GPI association had virtually identical kinetics. Antibody binding prolonged the exposure of the β2GPI/PS complex (t1/2: >1200min). The ability to phagocytose opsonized platelets was accordingly sustained (5.3±0.2 opsonized platelets were internalized by each macrophage after 60min and 9.4±0.3 after 300min). Anti-β2GPI antibodies therefore poise activated platelets in a PS-exposing status, preventing the recycling of their function and favoring their phagocytic clearance.