Ligation of Glycophorin A Generates Reactive Oxygen Species Leading to Decreased Red Blood Cell Function.
Authors:
Journal: PloS one
Publication Type: Journal Article
Date: 2016
DOI: PMC4718526
ID: 26784696
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.
Chemical List
- Anion Exchange Protein 1, Erythrocyte|||Glycophorins|||Reactive Oxygen Species|||Adenosine Triphosphate|||Caspase 3|||rac1 GTP-Binding Protein
Reference List
- Nelson RAJ (1953) The immune adherence phenomenon: an immunologically specfic reaction between microorganisms and erythrocytes leading to enhanced phagocytosis. Science 118: 733–737.|||Cooper NR (1969) Immune adherence by the fourth component of complement. Science 165: 396–398.|||Pisano A, Redmond JW, Williams KL, Gooley AA (1993) Glycosylation sites identified by solid-phase Edman degradation: O-linked glycosylation motifs on human glycophorin A. Glycobiology 3: 429–435.|||Abramson SB, Dobro J, Eberle MA, Benton M, Reibman J, et al. (1991) Acute reversible hypoxemia in systemic lupus erythematosus. Ann Int Med 114: 941–947.|||Ghiran IC, Zeidel ML, Shevkoplyas SS, Burns JM, Tsokos GC, et al. (2011) Systemic lupus erythematosus serum deposits C4d on red blood cells, decreases red blood cell membrane deformability, and promotes nitric oxide production. Arthritis and rheumatism 63: 503–512. 10.1002/art.30143|||Muroya T, Kannan L, Ghiran IC, Shevkoplyas SS, Paz Z, et al. (2014) C4d deposits on the surface of RBCs in trauma patients and interferes with their function. Crit Care Med 42: e364–372. 10.1097/CCM.0000000000000231|||Karnchanaphanurach P, Mirchev R, Ghiran I, Asara JM, Papahadjopoulos-Sternberg B, et al. (2009) C3b deposition on human erythrocytes induces the formation of a membrane skeleton-linked protein complex. J Clin Invest 119: 788–801. 10.1172/JCI36088|||Chasis JA, Mohandas N, Shohet SB (1985) Erythrocyte membrane rigidity induced by glycophorin A-ligand interaction. Evidence for a ligand-induced association between glycophorin A and skeletal proteins. J Clin Invest 75: 1919–1926.|||Brain MC, Prevost JM, Pihl CE, Brown CB (2002) Glycophorin A-mediated haemolysis of normal human erythrocytes: evidence for antigen aggregation in the pathogenesis of immune haemolysis. Br J Haematol 118: 899–908.|||De Backer D, Creteur J, Preiser JC, Dubois MJ, Vincent JL (2002) Microvascular blood flow is altered in patients with sepsis. Am J Respir Crit Care Med 166: 98–104.|||Orange JS, Hossny EM, Weiler CR, Ballow M, Berger M, et al. (2006) Use of intravenous immunoglobulin in human disease: a review of evidence by members of the Primary Immunodeficiency Committee of the American Academy of Allergy, Asthma and Immunology. J Allergy Clin Immunol 117: S525–553.|||Fruehauf JP, Meyskens FL Jr. (2007) Reactive oxygen species: a breath of life or death? Clin Cancer Res 13: 789–794.|||McMullin MF (1999) The molecular basis of disorders of the red cell membrane. J Clin Pathol 52: 245–248.|||Trautmann A (2009) Extracellular ATP in the immune system: more than just a "danger signal". Sci Signal 2: pe6 10.1126/scisignal.256pe6|||Matsuyama H, Amaya F, Hashimoto S, Ueno H, Beppu S, et al. (2008) Acute lung inflammation and ventilator-induced lung injury caused by ATP via the P2Y receptors: an experimental study. Respir Res 9: 79 10.1186/1465-9921-9-79|||Hasko G, Linden J, Cronstein B, Pacher P (2008) Adenosine receptors: therapeutic aspects for inflammatory and immune diseases. Nat Rev Drug Discov 7: 759–770. 10.1038/nrd2638|||Kolachala VL, Bajaj R, Chalasani M, Sitaraman SV (2008) Purinergic receptors in gastrointestinal inflammation. Am J Physiol Gastrointest Liver Physiol 294: G401–410.|||Inoue Y, Chen Y, Pauzenberger R, Hirsh MI, Junger WG (2008) Hypertonic saline up-regulates A3 adenosine receptor expression of activated neutrophils and increases acute lung injury after sepsis. Crit Care Med 36: 2569–2575. 10.1097/CCM.0b013e3181841a91|||Melhorn MI, Brodsky AS, Estanislau J, Khoory JA, Illigens B, et al. (2013) CR1-mediated ATP release by human red blood cells promotes CR1 clustering and modulates the immune transfer process. J Biol Chem 288: 31139–31153. 10.1074/jbc.M113.486035|||Bone RC, Balk RA, Cerra FB, Dellinger RP, Fein AM, et al. (2009) Definitions for sepsis and organ failure and guidelines for the use of innovative therapies in sepsis. The ACCP/SCCM Consensus Conference Committee. American College of Chest Physicians/Society of Critical Care Medicine. 1992. Chest 136: e28.|||Ghiran I, Glodek AM, Weaver G, Klickstein LB, Nicholson-Weller A (2008) Ligation of erythrocyte CR1 induces its clustering in complex with scaffolding protein FAP-1. Blood 112: 3465–3473. 10.1182/blood-2008-04-151845|||Pierce KR, Joyce JR, England RB, Jones LP (1972) Acute hemolytic anemia caused by wild onion poisoning in horses. J Am Vet Med Assoc 160: 323–327.|||Bloomfield CD, Brunning RD (1976) Acute leukemia as a terminal event in nonleukemic hematopoietic disorders. Sem in Oncol 3: 297–317.|||Ghiran IC, Zeidel ML, Shevkoplyas SS, Burns JM, Tsokos GC, et al. (2010) SLE serum deposits C4d on red blood cells, decreases red blood cell membrane deformability, and promotes nitric oxide production. Arthritis and rheumatism.|||Glodek AM, Mirchev R, Golan DE, Khoory JA, Burns JM, et al. (2010) Ligation of complement receptor 1 increases erythrocyte membrane deformability. Blood 116: 6063–6071. 10.1182/blood-2010-04-273904|||Vicas AE, Albrecht H, Lennox JL, del Rio C (2005) Imported malaria at an inner-city hospital in the United States. Am J Med Sci 329: 6–12.|||George A, Pushkaran S, Konstantinidis DG, Koochaki S, Malik P, et al. (2013) Erythrocyte NADPH oxidase activity modulated by Rac GTPases, PKC, and plasma cytokines contributes to oxidative stress in sickle cell disease. Blood 121: 2099–2107. 10.1182/blood-2012-07-441188|||Higuchi M, Honda T, Proske RJ, Yeh ET (1998) Regulation of reactive oxygen species-induced apoptosis and necrosis by caspase 3-like proteases. Oncogene 17: 2753–2760.|||Qu Y, Misaghi S, Newton K, Gilmour LL, Louie S, et al. (2011) Pannexin-1 is required for ATP release during apoptosis but not for inflammasome activation. J Immunol 186: 6553–6561. 10.4049/jimmunol.1100478|||Bao L, Locovei S, Dahl G (2004) Pannexin membrane channels are mechanosensitive conduits for ATP. FEBS Lett 572: 65–68.|||Abraham EH, Sterling KM, Kim RJ, Salikhova AY, Huffman HB, et al. (2001) Erythrocyte membrane ATP binding cassette (ABC) proteins: MRP1 and CFTR as well as CD39 (ecto-apyrase) involved in RBC ATP transport and elevated blood plasma ATP of cystic fibrosis. Blood cells, molecules & diseases 27: 165–180.|||Sprague RS, Ellsworth ML, Stephenson AH, Kleinhenz ME, Lonigro AJ (1998) Deformation-induced ATP release from red blood cells requires CFTR activity. Am J Physiol 275: H1726–1732.|||Harrison ML, Isaacson CC, Burg DL, Geahlen RL, Low PS (1994) Phosphorylation of human erythrocyte band 3 by endogenous p72syk. J Biol Chem 269: 955–959.|||Bohm R, Zaki L (1996) Towards the localization of the essential arginine residues in the band 3 protein of human red blood cell membranes. Biochim Biophys Acta 1280: 238–242.|||Moutzouri AG, Skoutelis AT, Gogos CA, Missirlis YF, Athanassiou GM (2007) Red blood cell deformability in patients with sepsis: a marker for prognosis and monitoring of severity. Clin Hemorheol Microcirc 36: 291–299.|||Hurd TC, Dasmahapatra KS, Rush BF Jr., Machiedo GW (1988) Red blood cell deformability in human and experimental sepsis. Arch Surg 123: 217–220.|||De Backer D, Orbegozo Cortes D, Donadello K, Vincent JL (2014) Pathophysiology of microcirculatory dysfunction and the pathogenesis of septic shock. Virulence 5: 73–79. 10.4161/viru.26482|||Zhou G, Kamenos G, Pendem S, Wilson JX, Wu F (2012) Ascorbate protects against vascular leakage in cecal ligation and puncture-induced septic peritonitis. Am J Physiol Regul Integr Comp Physiol 302: R409–416. 10.1152/ajpregu.00153.2011|||Miyano K, Sumimoto H (2007) Role of the small GTPase Rac in p22phox-dependent NADPH oxidases. Biochimie 89: 1133–1144.|||Berg CP, Engels IH, Rothbart A, Lauber K, Renz A, et al. (2001) Human mature red blood cells express caspase-3 and caspase-8, but are devoid of mitochondrial regulators of apoptosis. Cell death and differentiation 8: 1197–1206.|||Chu H, Low PS (2006) Mapping of glycolytic enzyme-binding sites on human erythrocyte band 3. Biochem J 400: 143–151.|||Betz T, Lenz M, Joanny JF, Sykes C (2009) ATP-dependent mechanics of red blood cells. Proc Natl Acad Sci U S A 106: 15320–15325. 10.1073/pnas.0904614106|||Park Y, Best CA, Auth T, Gov NS, Safran SA, et al. (2010) Metabolic remodeling of the human red blood cell membrane. Proc Natl Acad Sci U S A 107: 1289–1294. 10.1073/pnas.0910785107|||Wan J, Ristenpart WD, Stone HA (2008) Dynamics of shear-induced ATP release from red blood cells. Proc Natl Acad Sci U S A 105: 16432–16437. 10.1073/pnas.0805779105|||Borschukova O, Paz Z, Ghiran IC, Liu CC, Kao AH, et al. (2012) Complement fragment C3d is colocalized within the lipid rafts of T cells and promotes cytokine production. Lupus 21: 1294–1304.|||Bakhtiari N, Hosseinkhani S, Larijani B, Mohajeri-Tehrani MR, Fallah A (2012) Red blood cell ATP/ADP & nitric oxide: The best vasodilators in diabetic patients. J Diabetes Metab Disord 11: 9 10.1186/2251-6581-11-9|||Sinha A, Chu TT, Dao M, Chandramohanadas R (2015) Single-cell evaluation of red blood cell bio-mechanical and nano-structural alterations upon chemically induced oxidative stress. Sci Rep 5: 9768 10.1038/srep09768|||D'Alessandro A, Righetti PG, Zolla L (2010) The red blood cell proteome and interactome: an update. J Proteome Res 9: 144–163. 10.1021/pr900831f|||Brandao-Burch A, Key ML, Patel JJ, Arnett TR, Orriss IR (2012) The P2X7 Receptor is an Important Regulator of Extracellular ATP Levels. Front Endocrinol (Lausanne) 3: 41.|||Fan J, Zhang Y, Chuang-Smith ON, Frank KL, Guenther BD, et al. (2012) Ecto-5'-nucleotidase: a candidate virulence factor in Streptococcus sanguinis experimental endocarditis. PLoS One 7: e38059 10.1371/journal.pone.0038059|||MacFarlane GD, Sampson DE, Clawson DJ, Clawson CC, Kelly KL, et al. (1994) Evidence for an ecto-ATPase on the cell wall of Streptococcus sanguis. Oral Microbiol Immunol 9: 180–185.|||Wang L, Olivecrona G, Gotberg M, Olsson ML, Winzell MS, et al. (2005) ADP acting on P2Y13 receptors is a negative feedback pathway for ATP release from human red blood cells. Circ Res 96: 189–196.|||Martin C, Leone M, Viviand X, Ayem ML, Guieu R (2000) High adenosine plasma concentration as a prognostic index for outcome in patients with septic shock. Crit Care Med 28: 3198–3202.|||Chaplin H Jr., Coleman ME, Monroe MC (1983) In vivo instability of red-blood-cell-bound C3d and C4d. Blood 62: 965–971.|||Howell MD, Shapiro NI (2007) Surviving sepsis outside the intensive care unit. Crit Care Med 35: 1422–1423.|||Arnold RC, Shapiro NI, Jones AE, Schorr C, Pope J, et al. (2009) Multicenter study of early lactate clearance as a determinant of survival in patients with presumed sepsis. Shock 32: 35–39.|||Nencioni A, Trzeciak S, Shapiro NI (2009) The microcirculation as a diagnostic and therapeutic target in sepsis. Intern Emerg Med.|||Aird WC (2003) The role of the endothelium in severe sepsis and multiple organ dysfunction syndrome. Blood 101: 3765–3777.