Data from: Ligation of glycophorin A generates reactive oxygen species leading to decreased red blood cell function
Data files
Jan 21, 2016 version files 126.93 MB
-
Control C3D.004
32.77 KB
-
Control C3D.006
1 MB
-
control c3d.011
1 MB
-
control C3d.013
1 MB
-
Control C4D.005
65.54 KB
-
Control C4D.007
1 MB
-
control C4d.009
1 MB
-
control c4d.012
1 MB
-
control Ca 2+.006
1 MB
-
control DAF.005
1 MB
-
Control DAF.006
1 MB
-
Control DAF.008
1 MB
-
Control IgG.003
1 MB
-
Control IgG.005
1 MB
-
control IgG.007
1 MB
-
control IgG.009
1 MB
-
control NO.007
1 MB
-
control NO.010
1 MB
-
Control RBC + WGA.002
1 MB
-
control RBC + WGA.003
1 MB
-
Control RBC +IgG.005
1 MB
-
Control RBC.001
1 MB
-
Control RBC+C3d.007
1 MB
-
Control RBC+C4d.009
1 MB
-
Control RBC+DAF.011
1 MB
-
Control RBC+WGA.002
1 MB
-
Ctrl C3d
1 MB
-
Ctrl C4d
1 MB
-
Ctrl mIgG
1 MB
-
Ctrl RBC
1 MB
-
Ctrl WGA
1 MB
-
Data.001
1 MB
-
Data.002
1 MB
-
Data.003
1 MB
-
Data.004
1 MB
-
Data.005
1 MB
-
Data.006
1 MB
-
Data.007
1 MB
-
Data.008
1 MB
-
Data.009
1 MB
-
Data.010
1 MB
-
Data.011
1 MB
-
Data.012
1 MB
-
Flow Cytometry Data.xlsx
53.76 KB
-
M606 E0 + WGA.004
5.23 MB
-
M606 E0.003
5.15 MB
-
M606 E24 + WGA.002
5.67 MB
-
M606 E24.001
4.52 MB
-
M616E24 RBC+C4d.010
1 MB
-
M616E24 RBC+DAF.012
1 MB
-
M616E24.003
1 MB
-
M616E24+WGA.004
1 MB
-
M616E24RBC +IgG.006
1 MB
-
M616E24RBC+C3d.008
1 MB
-
M798E0 C3d
1 MB
-
M798E0 C4d
1 MB
-
M798E0 mIgG
1 MB
-
M798E0 RBC
1 MB
-
M798E0 WGA
1 MB
-
Normal Control RBC.001
1 MB
-
Normal Control RBC+WGA.002
1 MB
-
normal RBC + c3d.004
1 MB
-
normal RBC + c4d.005
1 MB
-
normal RBC + IgG.003
1 MB
-
normal RBC + WGA.002
1 MB
-
Sepsis +DAF.012
1 MB
-
sepsis c3d.008
1 MB
-
Sepsis C3D.010
1 MB
-
sepsis C3d.012
1 MB
-
sepsis C3d.014
1 MB
-
sepsis c4d.006
1 MB
-
sepsis C4d.010
1 MB
-
Sepsis C4D.011
1 MB
-
sepsis Ca 2+.005
1 MB
-
sepsis DAF.006
1 MB
-
sepsis IgG.005
1 MB
-
sepsis IgG.008
1 MB
-
Sepsis IgG.009
1 MB
-
sepsis IgG.010
1 MB
-
sepsis M616E0 RBC + c3d.009
1 MB
-
sepsis M616E0 RBC + c4d.010
1 MB
-
sepsis M616E0 RBC + IgG.008
1 MB
-
sepsis M616E0 RBC + WGA.007
1 MB
-
sepsis M616E0 RBC.006
1 MB
-
sepsis NO.007
1 MB
-
sepsis NO.008
1 MB
-
sepsis RBC + WGA.004
1 MB
-
Sepsis RBC C3D.010
1 MB
-
Sepsis RBC C4D.011
1 MB
-
Sepsis RBC DAF.012
1 MB
-
Sepsis RBC E0.007
1 MB
-
Sepsis RBC E0+WGA.008
1 MB
-
Sepsis RBC IgG.009
1 MB
-
sepsis RBC.002
1 MB
-
sepsis RBC.003
1 MB
-
Sepsis RBC+WGA.004
1 MB
-
Untreated normal RBC.001
1 MB
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.