Processing methods and storage duration impact extracellular vesicle counts in red blood cell units

Abstract

Extracellular vesicles (EVs) are active components of red blood cell (RBC) concentrates and may be associated with beneficial and adverse effects of transfusion. Elucidating controllable factors associated with EV release in RBC products is thus important to better manage the quality and properties of RBC units. Erythrocyte-derived EVs (EEVs) and platelet-derived EVs (PEVs) were counted in 1226 RBC units (administered to 280 patients) using a standardized cytometry-based method. EV size and CD47 and annexin V expression were also measured. The effects of donor characteristics, processing methods, and storage duration on EV counts were analyzed by using standard comparison tests, and analysis of covariance was used to determine factors independently associated with EV counts. PEV as well as EEV counts were higher in whole-blood–filtered RBC units compared with RBC-filtered units; PEV counts were associated with filter type (higher with filters associated with higher residual platelets), and CD47 expression was higher on EEVs in RBC units stored longer. Multivariate analysis showed that EEV counts were strongly associated with filter type (P < .0001), preparation, and storage time (+25.4 EEV/µL per day [P = .01] and +42.4 EEV/µL per day [P < .0001], respectively). The only independent factor associated with PEV counts was the residual platelet count in the unit (+67.1 PEV/µL; P < .0001). Overall, processing methods have an impact on EV counts and characteristics, leading to large variations in EV quantities transfused into patients. RBC unit processing methods might be standardized to control the EV content of RBC units if any impacts on patient outcomes can be confirmed. The IMIB (Impact of Microparticles in Blood) study is ancillary to the French ABLE (Age of Transfused Blood in Critically Ill Adults) trial (ISRCTN44878718).

Deformability of Stored Red Blood Cells

“…At the same time, it has been noted that the low temperature of cell storage stimulates the aggregation of rafts. Microvesicle counts in the RBCs bag are sensitive to the donor characteristics, unitprocessing methods, and storage duration (Almizraq et al, 2018;Gamonet et al, 2020).…”

Red Blood Cell Extracellular Vesicle-Based Drug Delivery: Challenges and Opportunities

“…RBCEVs are also enriched in synexin and sorcin, two proteins associated with stomatin-specific lipid rafts, as well as diacylglycerol and cholesterol as membrane lipids (24,44,48). Notably, the components of RBCEVs can be modified during RBC storage (47,55,56). Previous evidence illustrated that RBCEVs released from stored RBC units had increased surface CD47 expression and intravesicular miR-4454 and miR-451a levels over time (47,56).…”

“…Notably, the components of RBCEVs can be modified during RBC storage (47,55,56). Previous evidence illustrated that RBCEVs released from stored RBC units had increased surface CD47 expression and intravesicular miR-4454 and miR-451a levels over time (47,56). Concerning the membrane lipids, RBCEVs released after 4 weeks of RBC storage had higher ceramide, dihydroceramide, lysophosphatidylinositol, and lysophosphatidylglycerol levels, lower phosphatidylinositol and phosphatidylglycerol levels, but relatively unchanged phosphatidylethanolamine and lysophosphatidylethanolamine levels (55).…”

Extracellular Vesicles and Thrombosis: Update on the Clinical and Experimental Evidence

“…In addition to endothelial cells, platelets and white blood cells, EVs may also be generated from erythrocytes (red blood cells; RBCs), in particular from the membrane of aged or senescent erythrocytes [148]. RBC-derived EVs are found in high concentrations in erythrocyte concentrates, in particular after prolonged storage and following harsh processing methods [149], and are believed to be responsible for both beneficial and detrimental effects observed following blood transfusions [150]. Similar to platelets [137], they were shown to expose PS and bind lactadherin and annexin V and to assemble tenase and prothrombinase complex formation thus supporting thrombin generation, both in the presence and absence of added TF [151].…”