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Erythrocyte-Derived Microparticles Activate Pulmonary Endothelial Cells in a Murine Model of Transfusion

Chang, Alex L.; Kim, Young; Seitz, Aaron P.; Schuster, Rebecca M.; Lentsch, Alex B.; Pritts, Timothy A.

doi: 10.1097/SHK.0000000000000780
Basic Science Aspects
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ABSTRACT Erythrocyte-derived microparticles (MPs) are sub-micrometer, biologically active vesicles shed by red blood cells as part of the biochemical changes that occur during storage. We hypothesized that MPs from stored red blood cells would activate endothelial cells. MPs from aged murine packed red blood cells (pRBCs) were isolated and used to treat confluent layers of cultured endothelial cells. Endothelial expression of leukocyte adhesion molecules, endothelial-leukocyte adhesion molecule-1 (ELAM-1) and intercellular adhesion molecule-1(ICAM-1), and inflammatory mediator, interleukin-6 (IL-6), was evaluated at 0.5, 6, 12, and 24 h of treatment. Healthy C57BL/6 mice were transfused with a MP suspension and lung sections were analyzed for adhesion molecules and sequestered interstitial leukocytes. Increased levels of ELAM-1 and ICAM-1 were found on cultured endothelial cells 6 h after MP stimulation (6.91 vs. 4.07 relative fluorescent intensity [RFI], P < 0.01, and 5.85 vs. 3.55 RFI, P = 0.01, respectively). IL-6 in cell culture supernatants was increased after 12 h of MP stimulation compared with controls (1.24 vs. 0.73 ng/mL, P = 0.03). In vivo experiments demonstrated that MP injection increased ELAM-1 and ICAM-1 expression at 1 h (18.56 vs. 7.08 RFI, P < 0.01, and 23.66 vs. 6.87 RFI, P < 0.01, respectively) and caused increased density of pulmonary interstitial leukocytes by 4 h of treatment (69.25 vs. 29.25 cells/high powered field, P < 0.01). This series of experiments supports our hypothesis that erythrocyte-derived MPs are able to activate pulmonary endothelium, leading to the pulmonary sequestration of leukocytes following the transfusion of stored pRBCs.

Department of Surgery, University of Cincinnati, Cincinnati, Ohio

Address reprint requests to Timothy A. Pritts, MD, PhD, FACS, Associate Professor of Surgery, Department of Surgery, University of Cincinnati, 231 Albert Sabin Way, Mail Location 0558, Cincinnati, OH 45267-0558. E-mail:

Received 8 June, 2016

Revised 27 June, 2016

Accepted 19 October, 2016

Drs ALC and YK contributed equally to this work.

This work was supported by grants R01 GM107625 and T32 GM008478-23 from the National Institutes of Health.

The authors report no conflicts of interest.

© 2017 by the Shock Society