The Inflammatory Effects of Blood Donation-Derived Extracellular Microvesicles on a Monocytic THP-1 Cell Line Assessed by the Production of IL-8 and IL-10
Cardiff Metropolitan University
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Currently, contradictory evidence for Red Blood Cell (RBC) survival and mortality of patients post-transfusion of stored blood has led to an international disagreement over the clinical effects of transfusing stored blood. Platelet-derived microvesicles cause inflammation due to certain substrates expressed on their membranes. Extracellular microvesicles (EMVs) also contain these substrates and may play a role in the inflammatory response during transfusion of stored RBCs. This aims of this study were to determine the cell-of-origin of microvesicles derived from blood donation packs; and whether these microvesicles have inflammatory effects on THP-1 cells using IL-8 and IL-10 as indicators. Blood was extracted from SAG-M and Whole Blood packs donated from the Welsh Blood Service and centrifuged retaining the plasma or supernatant. EMVs were isolated using two centrifugation speeds, 17,000g and 100,000g, with flow cytometry and NanoSightTM performed to determine the cellular origin. THP-1 cells were incubated with EMVs and with or without LPS as an inflammatory inducer and the levels of IL-8 and IL-10 were measured using sandwich ELISA. The results of this study showed that EMVs derived from stored SAG-M packs were more homogenous and were in higher numbers than EMVs derived from fresh whole blood packs. EMVs derived from SAG-M blood packs at 17,000g elicited a pro- inflammatory effect on THP-1 cells without LPS stimulation, whereas separation at 100,000g did not show the same effect. EMVs derived from whole blood packs produced immunomodulatory effects shown by decreasing IL-8 production as the concentration of EMVs increased in the presence of LPS. Increases in the levels of IL-10 was shown when EMVs were present in low concentrations with LPS stimulation suggesting an immunomodulatory effect of EMVs at low concentrations. The conclusion of this study was that the transfusion of stored RBCs, compared to fresh RBCs, do not significantly affect post-transfusion mortality or morbidity of critically ill patients.
BSc (Hons) Biomedical Science
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