title: Storage of Erythrocytes Induces Suicidal Erythrocyte Death.
abstract: OBJECTIVE
  Similar to apoptosis of nucleated cells, red blood cells (RBC) can undergo suicidal cell death - called eryptosis. It is characterized by cell shrinkage and phosphatidylserine translocation. Eryptosis is triggered by an increase of intracellular calcium concentration due to activation of nonselective cation channels. The cation channels and consequently eryptosis are inhibited by erythropoietin. Eryptotic RBC are engulfed by macrophages and thus rapidly cleared from circulating blood. In this study, we explored whether storage of RBC influences the rate of eryptosis.
  
  
  METHODS
  Flow cytometry was employed to quantify phosphatidylserine exposing erythrocytes from annexin V binding and cytosolic Ca2+ activity from Fluo-3 fluorescence. Clearance of stored murine RBC was tested by injection of carboxyfluorescein succinimidyl ester (CFSE)-labelled erythrocytes.
  
  
  RESULTS
  Storage for 42 days significantly increased the percentage of phosphatidylserine exposing and haemolytic erythrocytes, an effect blunted by removal of extracellular calcium. Phosphatidylserine exposure could be inhibited by addition of erythropoietin. Upon transfusion, the clearance of murine CFSE-labelled RBC from circulating blood was significantly higher following storage for 10 days when compared to 2 days of storage.
  
  
  CONCLUSIONS
  Storage of RBC triggers eryptosis by Ca2+ and erythropoietin sensitive mechanisms.
authors.affiliation: Department of Gastroenterology, Hepatology and Infectiology, University of Dx00FC;sseldorf, Dx00FC;sseldorf, Germany.
authors.forename: Elisabeth
authors.initials: E
authors.lastname: Lang
authors.forename: Vitaly I
authors.initials: VI
authors.lastname: Pozdeev
authors.forename: Haifeng C
authors.initials: HC
authors.lastname: Xu
authors.forename: Prashant V
authors.initials: PV
authors.lastname: Shinde
authors.forename: Kristina
authors.initials: K
authors.lastname: Behnke
authors.forename: Junnat M
authors.initials: JM
authors.lastname: Hamdam
authors.forename: Erik
authors.initials: E
authors.lastname: Lehnert
authors.forename: Rüdiger E
authors.initials: RE
authors.lastname: Scharf
authors.forename: Florian
authors.initials: F
authors.lastname: Lang
authors.forename: Dieter
authors.initials: D
authors.lastname: Häussinger
authors.forename: Karl S
authors.initials: KS
authors.lastname: Lang
authors.forename: Philipp A
authors.initials: PA
authors.lastname: Lang
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects / Q000502:physiology*; D001692:Biological Transport / Q000187:drug effects; D001793:Blood Preservation / Q000379:methods*; D002118:Calcium / Q000378:metabolism; D016923:Cell Death / Q000187:drug effects / Q000502:physiology; D000072817:Eryptosis / Q000187:drug effects / Q000502:physiology*; D004912:Erythrocytes / Q000737:chemistry / Q000166:cytology / Q000378:metabolism*; D004921:Erythropoietin / Q000494:pharmacology; D005434:Flow Cytometry / Q000379:methods; D005452:Fluoresceins / Q000737:chemistry; D006801:Humans; D042541:Intracellular Space / Q000378:metabolism; D008810:Mice, Inbred C57BL; D010718:Phosphatidylserines / Q000378:metabolism; D013388:Succinimides / Q000737:chemistry; D013997:Time Factors
chemical_list: C087165:5-(6)-carboxyfluorescein diacetate succinimidyl ester; D005452:Fluoresceins; D010718:Phosphatidylserines; D013388:Succinimides; D004921:Erythropoietin; D002118:Calcium