Microparticles from stored red blood cells enhance procoagulant and proinflammatory activity

Abstract

BACKGROUND:The pathomechanisms of morbidity due to blood transfusions are not yet entirely understood. Elevated levels of red blood cell-derived microparticles (RMPs) are found in coagulation-related pathologies and also in stored blood. Previous research has shown that RMPs mediate transfusion-related complications by the intrinsic pathway. We hypothesized that RMPs might play a role in post-transfusion thrombotic complications by enhancing procoagulant activity also through the extrinsic pathway of coagulation. STUDY DESIGN AND METHODS:In this laboratory study, blood from 18 healthy volunteers was stimulated with microparticles from expired stored red blood cells. Various clotting parameters were recorded. Flow cytometry, enzyme-linked immunosorbent assays, and real-time polymerase chain reaction were used to investigate possible mediating mechanisms. RESULTS:The addition of RMPs shortened the clotting time from 194 to 161 seconds (p < 0.001). After incubation with RMPs, there was increased expression of tissue factor (TF) on monocytes and in plasma. TF messenger RNA expression increased in a timedependent and concentration-dependent manner. There was a significant induction of interleukin-1b and interleukin-6. After stimulation with RMPs, there was a significant increase in the number of activated platelets, an increased percentage of PAC-1/CD62P (procaspase activating compound-1/platelet surface P-selectin) double-positive platelets, and an increased number of platelet-neutrophil duplets and platelet-monocyte duplets, indicating enhanced interaction of platelets with neutrophils and monocytes. Levels of CXCL-8 (C-X-C motif chemokine ligand 1) and interleukin-6 were significantly higher after treatment with RMPs. CONCLUSION:Our results suggest that RMPs trigger coagulation through TF signaling, induce the secretion of proinflammatory cytokines, and induce cell-cell interaction between platelets and neutrophils. Thus, under certain conditions, RMPs could play a role in posttransfusion complications through these mechanisms. C ell-derived microparticles (MPs) are membrane vesicles smaller than 1 lm in diameter that are released by various types of cells during cell activation, injury, and apoptosis.1 Release ofMPs is also an integral part of red blood cell (RBC) aging and a prominent feature of the RBC storage lesion. It has been demonstrated that MPs are involved in a broad range of (patho-)physiological processes, such as angiogenesis, inflammation, and coagulation. Therefore, the biological effects and possible clinical relevance of RBCderived MPs (RMPs) need to be clarified. 2-5ABBREVIATION: CXCL-8 5 chemokine C-X-C motif ligand 8; Clinical studies have revealed that elevated levels of circulating RMPs are related to the hypercoagulable state in sickle-cell crisis, venous thromboembolism, and other conditions known to affect coagulation, such as diabetes, cardiovascular disease, and sepsis. [6][7][8][9][10] In addition, it has been observed that patients receiving RBCs are at risk for thrombotic events in a...

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