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PLoS One. 2016 Jan 19;11(1):e0141206. doi: 10.1371/journal.pone.0141206. eCollection 2016.

Ligation of Glycophorin A Generates Reactive Oxygen Species Leading to Decreased Red Blood Cell Function.

Author information

1
Division of Allergy and Inflammation, Beth Israel Deaconess Medical Center and Harvard Medical School, Boston, United States of America.
2
Department of Surgery, Brigham and Women's Hospital and Harvard Medical School, Boston, United States of America.
3
Department of Neurology, Beth Israel Deaconess Medical Center and Harvard Medical School, Boston, United States of America.
4
Department of Synthetic Chemistry & Biological Chemistry, Graduate School of Engineering, Kyoto University, Katsura, Kyoto, Japan.
5
The Barnett Institute of Chemical and Biological Analysis, Northeastern University, Boston, United States of America.
6
Department of Biomedical Engineering, Cullen College of Engineering, University of Houston, Houston, United States of America.
7
Department of Emergency Medicine and Center for Vascular Biology, Beth Israel Deaconess Medical Center and Harvard Medical School, Boston, United States of America.

Abstract

Acute, inflammatory conditions associated with dysregulated complement activation are characterized by significant increases in blood concentration of reactive oxygen species (ROS) and ATP. The mechanisms by which these molecules arise are not fully understood. In this study, using luminometric- and fluorescence-based methods, we show that ligation of glycophorin A (GPA) on human red blood cells (RBCs) results in a 2.1-fold, NADPH-oxidase-dependent increase in intracellular ROS that, in turn, trigger multiple downstream cascades leading to caspase-3 activation, ATP release, and increased band 3 phosphorylation. Functionally, using 2D microchannels to assess membrane deformability, GPS-ligated RBCs travel 33% slower than control RBCs, and lipid mobility was hindered by 10% using fluorescence recovery after photobleaching (FRAP). These outcomes were preventable by pretreating RBCs with cell-permeable ROS scavenger glutathione monoethyl ester (GSH-ME). Our results obtained in vitro using anti-GPA antibodies were validated using complement-altered RBCs isolated from control and septic patients. Our results suggest that during inflammatory conditions, circulating RBCs significantly contribute to capillary flow dysfunctions, and constitute an important but overlooked source of intravascular ROS and ATP, both critical mediators responsible for endothelial cell activation, microcirculation impairment, platelet activation, as well as long-term dysregulated adaptive and innate immune responses.

PMID:
26784696
PMCID:
PMC4718526
DOI:
10.1371/journal.pone.0141206
[Indexed for MEDLINE]
Free PMC Article

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