Density, heterogeneity and deformability of red cells as markers of clinical severity in hereditary spherocytosis
Rick Huisjes,
Asya Makhro,
Esther Llaudet-Planas,
Laura Hertz,
Polina Petkova-Kirova,
Liesbeth P. Verhagen,
Silvia Pignatelli,
Minke A.E. Rab,
Raymond M. Schiffelers,
Elena Seiler,
Wouter W. van Solinge,
Joan-LLuis Vives Corrons,
Lars Kaestner,
Maria Mañú-Pereira,
Anna Bogdanova,
Richard van Wijk
Affiliations
Rick Huisjes
Department of Clinical Chemistry and Hematology, University Medical Center Utrecht, Utrecht University, Utrecht, the Netherlands
Asya Makhro
Red Blood Cell Research Group, Institute of Veterinary Physiology, Vetsuisse Faculty and the Zurich Center for Integrative Human Physiology (ZIHP), University of Zurich, Zurich, Switzerland
Esther Llaudet-Planas
Red Blood Cell Defects and Hematopoietic Disorders Unit, Josep Carreras Leukemia Research Institute, Badalona, Barcelona, Spain
Laura Hertz
Theoretical Medicine and Biosciences, Medical Faculty, Saarland University, Homburg/Saar, Germany
Polina Petkova-Kirova
Theoretical Medicine and Biosciences, Medical Faculty, Saarland University, Homburg/Saar, Germany
Liesbeth P. Verhagen
Department of Clinical Chemistry and Hematology, University Medical Center Utrecht, Utrecht University, Utrecht, the Netherlands
Silvia Pignatelli
Department of Clinical Chemistry and Hematology, University Medical Center Utrecht, Utrecht University, Utrecht, the Netherlands
Minke A.E. Rab
Department of Clinical Chemistry and Hematology, University Medical Center Utrecht, Utrecht University, Utrecht, the Netherlands
Raymond M. Schiffelers
Department of Clinical Chemistry and Hematology, University Medical Center Utrecht, Utrecht University, Utrecht, the Netherlands
Elena Seiler
Red Blood Cell Research Group, Institute of Veterinary Physiology, Vetsuisse Faculty and the Zurich Center for Integrative Human Physiology (ZIHP), University of Zurich, Zurich, Switzerland
Wouter W. van Solinge
Department of Clinical Chemistry and Hematology, University Medical Center Utrecht, Utrecht University, Utrecht, the Netherlands
Joan-LLuis Vives Corrons
Red Blood Cell Defects and Hematopoietic Disorders Unit, Josep Carreras Leukemia Research Institute, Badalona, Barcelona, Spain
Lars Kaestner
Theoretical Medicine and Biosciences, Medical Faculty, Saarland University, Homburg/Saar, Germany;Experimental Physics, Saarland University, Saarbruecken, Germany
Maria Mañú-Pereira
Rare Anemia Research Unit. Vall d’Hebron Research Institution, University Hospital Vall d’Hebron, Barcelona, Spain
Anna Bogdanova
Red Blood Cell Research Group, Institute of Veterinary Physiology, Vetsuisse Faculty and the Zurich Center for Integrative Human Physiology (ZIHP), University of Zurich, Zurich, Switzerland
Richard van Wijk
Department of Clinical Chemistry and Hematology, University Medical Center Utrecht, Utrecht University, Utrecht, the Netherlands
Hereditary spherocytosis (HS) originates from defective anchoring of the cytoskeletal network to the transmembrane protein complexes of the red blood cell (RBC). Red cells in HS are characterized by membrane instability and reduced deformability and there is marked heterogeneity in disease severity among patients. To unravel this variability in disease severity, we analyzed blood samples from 21 HS patients with defects in ankyrin, band 3, α-spectrin or β-spectrin using red cell indices, eosin-5-maleimide binding, microscopy, the osmotic fragility test, Percoll density gradients, vesiculation and ektacytometry to assess cell membrane stability, cellular density and deformability. Reticulocyte counts, CD71 abundance, band 4.1 a:b ratio, and glycated hemoglobin were used as markers of RBC turnover. We observed that patients with moderate/severe spherocytosis have short-living erythrocytes of low density and abnormally high intercellular heterogeneity. These cells show a prominent decrease in membrane stability and deformability and, as a consequence, are quickly removed from the circulation by the spleen. In contrast, in mild spherocytosis less pronounced reduction in deformability results in prolonged RBC lifespan and, hence, cells are subject to progressive loss of membrane. RBC from patients with mild spherocytosis thus become denser before they are taken up by the spleen. Based on our findings, we conclude that RBC membrane loss, cellular heterogeneity and density are strong markers of clinical severity in spherocytosis.
