Suicidal death of erythrocytes in recurrent hemolytic uremic syndrome

  • Philipp A. Lang
  • Ortraud Beringer
  • Jan P. Nicolay
  • Oliver Amon
  • Daniela S. Kempe
  • Tobias Hermle
  • Philipp Attanasio
  • Ahmad Akel
  • Richard Schäfer
  • Björn Friedrich
  • Teut Risler
  • Matthias Baur
  • Christoph J. Olbricht
  • Lothar Bernd Zimmerhackl
  • Peter F. Zipfel
  • Thomas Wieder
  • Florian Lang
Original Article

DOI: 10.1007/s00109-006-0058-0

Cite this article as:
Lang, P.A., Beringer, O., Nicolay, J.P. et al. J Mol Med (2006) 84: 378. doi:10.1007/s00109-006-0058-0

Abstract

Hemolytic uremic syndrome (HUS) is characterized by hemolytic anemia with fragmented erythrocytes, thrombocytopenia, and acute renal failure. Lack of complement inactivating factor H predisposes to the development of atypical HUS. Little is known about mechanisms linking complement activation with loss of erythrocyte integrity during HUS. Recent studies disclosed that increased cytosolic Ca2+ activity and cellular ceramide trigger programmed erythrocyte death or eryptosis, characterized by cell shrinkage and phosphatidylserine exposure at the erythrocyte surface. In the present study, we investigated whether eryptosis occurs during the course of HUS. To this end, erythrocytes from healthy volunteers were exposed to plasma from a patient with severe idiopathic recurrent HUS secondary to factor H depletion. Phosphatidylserine exposure (Annexin binding), cell volume (forward scatter), cytosolic Ca2+ activity (Fluo3 fluorescence), and ceramide formation [anti-ceramide antibody and enzymatic (diacylgycerol kinase) analysis] were determined. Exposure of erythrocytes to plasma from the patient, but not to plasma from healthy individuals, triggered Annexin binding. The effect of plasma on erythrocyte Annexin binding was abolished by plasmapheresis or filtration at 30 kDa. It was paralleled by formation of ceramide and increase of cytosolic Ca2+ activity. Enhanced Annexin binding of erythrocytes from healthy individuals was observed after exposure to plasma from three other patients with HUS. The proeryptotic effect of patient plasma was mimicked by exposure to the Ca2+ ionophore ionomycin, and eryptosis was potentiated in the presence of cell membrane-permeable C6-ceramide. Furthermore, in vitro complement activation similarly triggered erythrocyte phosphatidylserine exposure, an effect which was blunted by the addition of factor H. In conclusion, our present observations disclose a novel, pathophysiological, factor-H dependent mechanism leading to injury of erythrocytes during the course of hemolytic uremic syndrome.

Keywords

ApoptosisPhosphatidylserineSphingomyelinaseCeramideCalciumPlasmapheresis

Copyright information

© Springer-Verlag 2006

Authors and Affiliations

  • Philipp A. Lang
    • 1
  • Ortraud Beringer
    • 2
  • Jan P. Nicolay
    • 1
  • Oliver Amon
    • 2
  • Daniela S. Kempe
    • 1
  • Tobias Hermle
    • 1
  • Philipp Attanasio
    • 1
  • Ahmad Akel
    • 1
  • Richard Schäfer
    • 3
  • Björn Friedrich
    • 4
  • Teut Risler
    • 4
  • Matthias Baur
    • 5
  • Christoph J. Olbricht
    • 5
  • Lothar Bernd Zimmerhackl
    • 6
  • Peter F. Zipfel
    • 7
  • Thomas Wieder
    • 1
  • Florian Lang
    • 1
  1. 1.Department of PhysiologyUniversity of TübingenTübingenGermany
  2. 2.Children’s University Hospital TübingenTübingenGermany
  3. 3.Department of Transfusion MedicineUniversity Hospital TübingenTübingenGermany
  4. 4.Department of Internal MedicineUniversity Hospital TübingenTübingenGermany
  5. 5.KatharinenhospitalStuttgartGermany
  6. 6.Department of PediatricsUniversity of InnsbruckInnsbruckAustria
  7. 7.Department of Biological InfectionHans Knoell Institute for Natural Products ResearchJenaGermany