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Suicidal death of erythrocytes in recurrent hemolytic uremic syndrome

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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.

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Acknowledgements

This study was supported by the Deutsche Forschungsgemeinschaft, Nr. La 315/4-3, La 315/6-1, and La 315/13-1. P. A. L., D. K., and J. N. were recipients of a stipend from the Center for Interdisciplinary Clinical Research, IZKF (Fö. 01KS9602; Promotionskolleg “Molekulare Medizin # 1547”).

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Authors and Affiliations

  1. Department of Physiology, University of Tübingen, Gmelinstr. 5, 72076, Tübingen, Germany

    Philipp A. Lang, Jan P. Nicolay, Daniela S. Kempe, Tobias Hermle, Philipp Attanasio, Ahmad Akel, Thomas Wieder & Florian Lang

  2. Children’s University Hospital Tübingen, Hoppe Seyler Str. 1, 72076, Tübingen, Germany

    Ortraud Beringer & Oliver Amon

  3. Department of Transfusion Medicine, University Hospital Tübingen, Hoppe Seyler Str. 1, 72076, Tübingen, Germany

    Richard Schäfer

  4. Department of Internal Medicine, University Hospital Tübingen, Hoppe Seyler Str. 1, 72076, Tübingen, Germany

    Björn Friedrich & Teut Risler

  5. Katharinenhospital, Kriegsbergstr. 60, 70174, Stuttgart, Germany

    Matthias Baur & Christoph J. Olbricht

  6. Department of Pediatrics, University of Innsbruck, Anichstr. 35, A6020, Innsbruck, Austria

    Lothar Bernd Zimmerhackl

  7. Department of Biological Infection, Hans Knoell Institute for Natural Products Research, Beutenbergstr.11a, 07745, Jena, Germany

    Peter F. Zipfel

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  1. Philipp A. Lang
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  2. Ortraud Beringer
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Lang, P.A., Beringer, O., Nicolay, J.P. et al. Suicidal death of erythrocytes in recurrent hemolytic uremic syndrome. J Mol Med 84, 378–388 (2006). https://doi.org/10.1007/s00109-006-0058-0

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