DOI: 10.5061/dryad.f862c
title: Data from: Ligation of glycophorin A generates reactive oxygen species leading to decreased red blood cell function

description: 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.
description: Flow Cytometry Data of
 Sepsis StudyFlow cytometry data
 including C3d, C4d, glycocalyx (WGA), and some other data (FRAP,
 etc.)Flow
 Cytometry Data.xlsxM606 E0 +
 WGAFlow cytometry raw data.
 Flow cytometry files (.fcs) generated using a LSRII flow
 cytometer.M606
 E0Flow cytometry raw data.
 Flow cytometry files (.fcs) generated using a LSRII flow
 cytometer.M606 E24 +
 WGAFlow cytometry raw data.
 Flow cytometry files (.fcs) generated using a LSRII flow
 cytometer.M606
 E24Flow cytometry raw data.
 Flow cytometry files (.fcs) generated using a LSRII flow
 cytometer.M613E24-control
 C3dFlow cytometry raw data.
 Flow cytometry files (.fcs) generated using a LSRII flow
 cytometer.control
 C3d.011M613E24-control
 C4dFlow cytometry raw data.
 Flow cytometry files (.fcs) generated using a LSRII flow
 cytometer.control
 C4d.009M613E24-control
 DAFFlow cytometry raw data.
 Flow cytometry files (.fcs) generated using a LSRII flow
 cytometer.control
 DAF.005M613E24-control
 IgGFlow cytometry raw data.
 Flow cytometry files (.fcs) generated using a LSRII flow
 cytometer.control
 IgG.007M613E24-control RBC +
 WGAFlow cytometry raw data.
 Flow cytometry files (.fcs) generated using a LSRII flow
 cytometer.control RBC +
 WGA.002M613E24-control
 RBCFlow cytometry raw data.
 Flow cytometry files (.fcs) generated using a LSRII flow
 cytometer.control
 RBC.001M613E24-sepsis
 C3dFlow cytometry raw data.
 Flow cytometry files (.fcs) generated using a LSRII flow
 cytometer.sepsis
 C3d.012M613E24-sepsis
 C4dFlow cytometry raw data.
 Flow cytometry files (.fcs) generated using a LSRII flow
 cytometer.sepsis
 C4d.010M613E24-sepsis
 DAFFlow cytometry raw data.
 Flow cytometry files (.fcs) generated using a LSRII flow
 cytometer.sepsis
 DAF.006M613E24-sepsis
 IgGFlow cytometry raw data.
 Flow cytometry files (.fcs) generated using a LSRII flow
 cytometer.sepsis
 IgG.008M613E24-sepsis RBC +
 WGAFlow cytometry raw data.
 Flow cytometry files (.fcs) generated using a LSRII flow
 cytometer.sepsis RBC +
 WGA.004M613E24-sepsis
 RBCFlow cytometry raw data.
 Flow cytometry files (.fcs) generated using a LSRII flow
 cytometer.sepsis
 RBC.003M613E48-control
 C3dFlow cytometry raw data.
 Flow cytometry files (.fcs) generated using a LSRII flow
 cytometer.control
 C3d.013M613E48-control
 C4dFlow cytometry raw data.
 Flow cytometry files (.fcs) generated using a LSRII flow
 cytometer.control
 C4d.012M613E48-control Ca
 2+Flow cytometry raw data.
 Flow cytometry files (.fcs) generated using a LSRII flow
 cytometer.control Ca
 2+.006M613E48-control
 IgGFlow cytometry raw data.
 Flow cytometry files (.fcs) generated using a LSRII flow
 cytometer.control
 IgG.009M613E48-control
 NOFlow cytometry raw data.
 Flow cytometry files (.fcs) generated using a LSRII flow
 cytometer.control
 NO.007M613E48-control RBC +
 WGAFlow cytometry raw data.
 Flow cytometry files (.fcs) generated using a LSRII flow
 cytometer.control RBC +
 WGA.003M613E48-control
 RBCFlow cytometry raw data.
 Flow cytometry files (.fcs) generated using a LSRII flow
 cytometer.control
 RBC.001M613E48-sepsis
 C3dFlow cytometry raw data.
 Flow cytometry files (.fcs) generated using a LSRII flow
 cytometer.sepsis
 C3d.014M613E48-sepsis
 C4dFlow cytometry raw data.
 Flow cytometry files (.fcs) generated using a LSRII flow
 cytometer.sepsis
 C4d.011M613E48-sepsis Ca
 2+Flow cytometry raw data.
 Flow cytometry files (.fcs) generated using a LSRII flow
 cytometer.sepsis Ca
 2+.005M613E48-sepsis
 IgGFlow cytometry raw data.
 Flow cytometry files (.fcs) generated using a LSRII flow
 cytometer.sepsis
 IgG.010M613E48-sepsis
 NOFlow cytometry raw data.
 Flow cytometry files (.fcs) generated using a LSRII flow
 cytometer.sepsis
 NO.008M613E48-sepsis RBC +
 WGAFlow cytometry raw data.
 Flow cytometry files (.fcs) generated using a LSRII flow
 cytometer.sepsis RBC +
 WGA.004M613E48-sepsis
 RBCFlow cytometry raw data.
 Flow cytometry files (.fcs) generated using a LSRII flow
 cytometer.sepsis
 RBC.002M616E0-normal RBC +
 c3dFlow cytometry raw data.
 Flow cytometry files (.fcs) generated using a LSRII flow
 cytometer.normal RBC +
 c3d.004M616E0-normal RBC +
 c4dFlow cytometry raw data.
 Flow cytometry files (.fcs) generated using a LSRII flow
 cytometer.normal RBC +
 c4d.005M616E0-normal RBC +
 IgGFlow cytometry raw data.
 Flow cytometry files (.fcs) generated using a LSRII flow
 cytometer.normal RBC +
 IgG.003M616E0-normal RBC +
 WGAFlow cytometry raw data.
 Flow cytometry files (.fcs) generated using a LSRII flow
 cytometer.normal RBC +
 WGA.002M616E0-Untreated normal
 RBCFlow cytometry raw data.
 Flow cytometry files (.fcs) generated using a LSRII flow
 cytometer.Untreated normal
 RBC.001M616E0 RBC +
 c3dFlow cytometry raw data.
 Flow cytometry files (.fcs) generated using a LSRII flow
 cytometer.sepsis M616E0 RBC +
 c3d.009M616E0 RBC +
 c4dFlow cytometry raw data.
 Flow cytometry files (.fcs) generated using a LSRII flow
 cytometer.sepsis M616E0 RBC +
 c4d.010M616E0 RBC +
 IgGFlow cytometry raw data.
 Flow cytometry files (.fcs) generated using a LSRII flow
 cytometer.sepsis M616E0 RBC +
 IgG.008M616E0 RBC +
 WGAFlow cytometry raw data.
 Flow cytometry files (.fcs) generated using a LSRII flow
 cytometer.sepsis M616E0 RBC +
 WGA.007M616E0
 RBCFlow cytometry raw data.
 Flow cytometry files (.fcs) generated using a LSRII flow
 cytometer.sepsis M616E0
 RBC.006M616E24-Control RBC
 +IgGFlow cytometry raw data.
 Flow cytometry files (.fcs) generated using a LSRII flow
 cytometer.Control RBC
 +IgG.005M616E24-Control
 RBC+C3dFlow cytometry raw data.
 Flow cytometry files (.fcs) generated using a LSRII flow
 cytometer.Control
 RBC+C3d.007M616E24-Control
 RBC+C4dFlow cytometry raw data.
 Flow cytometry files (.fcs) generated using a LSRII flow
 cytometer.Control
 RBC+C4d.009M616E24-Control
 RBC+DAFFlow cytometry raw data.
 Flow cytometry files (.fcs) generated using a LSRII flow
 cytometer.Control
 RBC+DAF.011M616E24
 RBC+C4dFlow cytometry raw data.
 Flow cytometry files (.fcs) generated using a LSRII flow
 cytometer.M616E24
 RBC+DAFFlow cytometry raw data.
 Flow cytometry files (.fcs) generated using a LSRII flow
 cytometer.M616E24
 RBCFlow cytometry raw data.
 Flow cytometry files (.fcs) generated using a LSRII flow
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Flow cytometry files (.fcs) generated using a LSRII flow cytometer.Control DAF.008M618E0-Control IgGFlow cytometry raw data. Flow cytometry files (.fcs) generated using a LSRII flow cytometer.Control IgG.005M618E0-Control RBCFlow cytometry raw data. Flow cytometry files (.fcs) generated using a LSRII flow cytometer.Control RBC.001M618E0-Control RBC+WGAFlow cytometry raw data. Flow cytometry files (.fcs) generated using a LSRII flow cytometer.Control RBC+WGA.002M618E0-Sepsis +DAFFlow cytometry raw data. Flow cytometry files (.fcs) generated using a LSRII flow cytometer.Sepsis +DAF.012M618E0-Sepsis C3DFlow cytometry raw data. Flow cytometry files (.fcs) generated using a LSRII flow cytometer.Sepsis C3D.010M618E0-Sepsis C4DFlow cytometry raw data. Flow cytometry files (.fcs) generated using a LSRII flow cytometer.Sepsis C4D.011M618E0-Sepsis IgGFlow cytometry raw data. Flow cytometry files (.fcs) generated using a LSRII flow cytometer.Sepsis IgG.009M618E0-Sepsis RBCFlow cytometry raw data. Flow cytometry files (.fcs) generated using a LSRII flow cytometer.Sepsis RBC.003M618E0-Sepsis RBC+WGAFlow cytometry raw data. Flow cytometry files (.fcs) generated using a LSRII flow cytometer.Sepsis RBC+WGA.004M618E24-Data1Flow cytometry raw data. Flow cytometry files (.fcs) generated using a LSRII flow cytometer.Data.001M618E24-Data2Flow cytometry raw data. Flow cytometry files (.fcs) generated using a LSRII flow cytometer.Data.002M618E24-Data3Flow cytometry raw data. Flow cytometry files (.fcs) generated using a LSRII flow cytometer.Data.003M618E24-Data4Flow cytometry raw data. Flow cytometry files (.fcs) generated using a LSRII flow cytometer.Data.004M618E24-Data5Flow cytometry raw data. Flow cytometry files (.fcs) generated using a LSRII flow cytometer.Data.005M618E24-Data6Flow cytometry raw data. Flow cytometry files (.fcs) generated using a LSRII flow cytometer.Data.006M618E24-Data7Flow cytometry raw data. Flow cytometry files (.fcs) generated using a LSRII flow cytometer.Data.007M618E24-Data8Flow cytometry raw data. Flow cytometry files (.fcs) generated using a LSRII flow cytometer.Data.008M618E24-Data9Flow cytometry raw data. Flow cytometry files (.fcs) generated using a LSRII flow cytometer.Data.009M618E24-Data10Flow cytometry raw data. Flow cytometry files (.fcs) generated using a LSRII flow cytometer.Data.010M618E24-Data11Flow cytometry raw data. Flow cytometry files (.fcs) generated using a LSRII flow cytometer.Data.011M618E24-Data12Flow cytometry raw data. Flow cytometry files (.fcs) generated using a LSRII flow cytometer.Data.012M625E0-Control C3DFlow cytometry raw data. Flow cytometry files (.fcs) generated using a LSRII flow cytometer.Control C3D.004M625E0-Control C4DFlow cytometry raw data. Flow cytometry files (.fcs) generated using a LSRII flow cytometer.Control C4D.005M625E0-Control DAFFlow cytometry raw data. Flow cytometry files (.fcs) generated using a LSRII flow cytometer.Control DAF.006M625E0-Control IgGFlow cytometry raw data. Flow cytometry files (.fcs) generated using a LSRII flow cytometer.Control IgG.003M625E0-Control RBCFlow cytometry raw data. Flow cytometry files (.fcs) generated using a LSRII flow cytometer.Control RBC.001M625E0-Control RBC+WGAFlow cytometry raw data. Flow cytometry files (.fcs) generated using a LSRII flow cytometer.Control RBC+WGA.002M625E0-Sepsis RBC C3DFlow cytometry raw data. Flow cytometry files (.fcs) generated using a LSRII flow cytometer.Sepsis RBC C3D.010M625E0-Sepsis RBC C4DFlow cytometry raw data. Flow cytometry files (.fcs) generated using a LSRII flow cytometer.Sepsis RBC C4D.011M625E0-Sepsis RBC DAFFlow cytometry raw data. Flow cytometry files (.fcs) generated using a LSRII flow cytometer.Sepsis RBC DAF.012M625E0-Sepsis RBC IgGFlow cytometry raw data. Flow cytometry files (.fcs) generated using a LSRII flow cytometer.Sepsis RBC IgG.009M625E0-Sepsis RBC E0Flow cytometry raw data. Flow cytometry files (.fcs) generated using a LSRII flow cytometer.Sepsis RBC E0.007M625E0-Sepsis RBC E0+WGAFlow cytometry raw data. Flow cytometry files (.fcs) generated using a LSRII flow cytometer.Sepsis RBC E0+WGA.008M625E24-control c3dFlow cytometry raw data. Flow cytometry files (.fcs) generated using a LSRII flow cytometer.control c3d.011M625E24-control c4dFlow cytometry raw data. Flow cytometry files (.fcs) generated using a LSRII flow cytometer.control c4d.012M625E24-control IgGFlow cytometry raw data. Flow cytometry files (.fcs) generated using a LSRII flow cytometer.control IgG.009M625E24-control NOFlow cytometry raw data. Flow cytometry files (.fcs) generated using a LSRII flow cytometer.control NO.010M625E24-Control RBC + WGAFlow cytometry raw data. Flow cytometry files (.fcs) generated using a LSRII flow cytometer.Control RBC + WGA.002M625E24-Control RBCFlow cytometry raw data. Flow cytometry files (.fcs) generated using a LSRII flow cytometer.Control RBC.001M625E24-sepsis c3dFlow cytometry raw data. Flow cytometry files (.fcs) generated using a LSRII flow cytometer.sepsis c3d.008M625E24-sepsis c4dFlow cytometry raw data. Flow cytometry files (.fcs) generated using a LSRII flow cytometer.sepsis c4d.006M625E24-sepsis NOFlow cytometry raw data. Flow cytometry files (.fcs) generated using a LSRII flow cytometer.sepsis NO.007M625E24-sepsis RBC + WGAFlow cytometry raw data. Flow cytometry files (.fcs) generated using a LSRII flow cytometer.sepsis RBC + WGA.004M625E24-sepsis RBCFlow cytometry raw data. Flow cytometry files (.fcs) generated using a LSRII flow cytometer.sepsis RBC.003M625E24-sepsis IgGFlow cytometry raw data. Flow cytometry files (.fcs) generated using a LSRII flow cytometer.sepsis IgG.005M630E24-Control C3DFlow cytometry raw data. Flow cytometry files (.fcs) generated using a LSRII flow cytometer.Control C3D.004M630E24-Control C4DFlow cytometry raw data. Flow cytometry files (.fcs) generated using a LSRII flow cytometer.Control C4D.005M798E0 Ctrl C3DFlow cytometry raw data. Flow cytometry files (.fcs) generated using a LSRII flow cytometer.Ctrl C3dM798E0 Ctrl C4DFlow cytometry raw data. Flow cytometry files (.fcs) generated using a LSRII flow cytometer.Ctrl C4dM798E0 Ctrl mIgGFlow cytometry raw data. Flow cytometry files (.fcs) generated using a LSRII flow cytometer.Ctrl mIgGM798E0 Ctrl RBCFlow cytometry raw data. Flow cytometry files (.fcs) generated using a LSRII flow cytometer.Ctrl RBCM798E0 Ctrl WGAFlow cytometry raw data. Flow cytometry files (.fcs) generated using a LSRII flow cytometer.Ctrl WGAM798E0 C3DFlow cytometry raw data. Flow cytometry files (.fcs) generated using a LSRII flow cytometer.M798E0 C3dM798E0 C4DFlow cytometry raw data. Flow cytometry files (.fcs) generated using a LSRII flow cytometer.M798E0 C4dM798E0 mIgGFlow cytometry raw data. Flow cytometry files (.fcs) generated using a LSRII flow cytometer.M798E0 RBCFlow cytometry raw data. Flow cytometry files (.fcs) generated using a LSRII flow cytometer.M798E0 WGAFlow cytometry raw data. Flow cytometry files (.fcs) generated using a LSRII flow cytometer.