Supernatants and lipids from stored red blood cells activate pulmonary microvascular endothelium through the BLT2 receptor and protein kinase C activation

• Published in: Transfusion (Philadelphia, Pa.) Vol. 57; no. 11; pp. 2690 - 2700
• Main Authors: Silliman, Christopher C., Kelher, Marguerite R., Khan, Samina Y., West, F. Bernadette, McLaughlin, Nathan J.D., Elzi, David J., England, Kelly, Bjornsen, Jason, Kuldanek, Susan A., Banerjee, Anirban
• Format: Journal Article
• Language: English
• Published: United States Wiley Subscription Services, Inc 01-11-2017
• Subjects: Acute Lung Injury - etiology; Adhesion; Adhesion tests; Arrestin; Blood Preservation - methods; Cell activation; Cell adhesion; Culture Media, Conditioned - pharmacology; Endothelial cells; Endothelial Cells - metabolism; Endothelium; Enzyme Activation; Erythrocyte Transfusion - adverse effects; Erythrocytes; Erythrocytes - cytology; Humans; Inflammation; Injuries; Intercellular adhesion molecule 1; Interleukin 8; Kinases; Lipids; Lipids - pharmacology; Lung - blood supply; Lungs; Microvasculature; Microvessels - cytology; Morbidity; Patients; Pneumonia - etiology; Protein kinase C; Protein Kinase C - metabolism; Receptors, Leukotriene B4 - metabolism; Signaling; Storage; Transfusion
• ISSN: 0041-1132; 1537-2995
• DOI: 10.1111/trf.14271

BACKGROUND
Although transfusion is a lifesaving intervention, it may be associated with significant morbidity in injured patients. We hypothesize that stored red blood cells (RBCs) induce proinflammatory activation of human pulmonary microvascular endothelial cells (HMVECs) resulting in neutrophil (PMN) adhesion and predisposition to acute lung injury (ALI).
STUDY DESIGN AND METHODS
Ten units of RBCs were collected; 50% (by weight) were leukoreduced (LR‐RBCs) and the remainder was unmodified and stored in additive solution‐5 (AS‐5). An additional 10 units of RBCs were collected, leukoreduced, and stored in AS‐3. HMVECs were incubated with [10%‐40%]FINAL of the supernatants on Day (D)1 to D42 of storage, lipid extracts, and purified lipids. Endothelial surface expression of intercellular adhesion molecule‐1 (ICAM‐1), interleukin (IL)‐8 release, and PMN adhesion to HMVECs were measured. HMVEC signaling via the BLT2 receptor was evaluated. Supernatants and lipids were also employed as the first event in a two‐event model of ALI.
RESULTS
The supernatants [10%‐40%]FINAL from D21 LR‐RBCs and D42 RBCs and LR‐RBCs and the lipids from D42 stored in AS‐5 induced increased ICAM‐1 surface expression on endothelium, IL‐8 release, and PMN adhesion. In addition, the supernatants [20%‐40%]FINAL from D21 and D42 RBCs in AS‐5 also increased endothelial surface expression of ICAM‐1. D42 supernatants and lipids also caused coprecipitation of β‐arrestin‐1 with BLT2, protein kinase C (PKC)βI, and PKCδ and served as the first event in a two‐event rodent model of ALI.
CONCLUSION
Lipids that accumulate during RBC storage activate endothelium and predispose to ALI, which may explain some of the adverse events associated with the transfusion of critically injured patients.