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Prepublished online as a Blood First Edition Paper on February 27, 2003; DOI 10.1182/blood-2002-11-3416.
Submitted November 14, 2002
Accepted February 13, 2003
Phosphatidylserine externalization in sickle red blood cells: associations with cell age, density, and hemoglobin F
Zahida Yasin*, Scott Witting, Mary B Palascak, Clinton H Joiner, Donald L Rucknagel, and Robert S Franco
Department of Internal Medicine, University of Cincinnati College of Medicine, Cincinnati, OH, USA
Department of Pathology and Laboratory Medicine, University of Cincinnati College of Medicine, Cincinnati, OH, USA
Department of Pediatrics, Cincinnati Comprehensive Sickle Cell Center, Cincinnati, OH, USA
Department of Pediatrics, Children's Hospital Research Foundation, Cincinnati, OH, USA
* Corresponding author; email: yasinz{at}email.uc.edu.
Phosphatidylserine (PS) is normally confined to the cytoplasmic leaflet of the red blood cell (RBC) membrane, but some sickle RBC expose PS in the outer leaflet (PS+ cells). This study examined the relationships among PS externalization, fetal hemoglobin content, hydration state, and cell age. Sickle RBC exhibit a wide range of PS externalization. Those with low level exposure (Type I PS+) include many young transferrin receptor positive (TfR+) cells. This is not specific for sickle cell disease, since many non-sickle TfR+ cells were also PS+. Sickle RBC with higher PS exposure (Type II PS+) appear to be more specific for sickle cell disease. Their formation is most likely sickling dependent, since Type II PS+ dense sickle cells had a lower percent HbF than PS- cells in the same density fraction (1.7 vs. 2.9, n=8, P < 0.01). In vivo experiments using biotin-labeled sickle cells showed a sharp decrease in the percentage of circulating, labeled PS+ cells during the first twenty-four hours after reinfusion. This decrease was confined to Type I PS+ cells, and was thus consistent with reversal of PS exposure in very young cells. As the labeled cells aged in the circulation, the percentage of both Type I and Type II PS+ cells increased. These studies indicate that PS externalization in sickle cells may be low level, as observed in many immature cells, or high level, which is associated with dehydration and appears to be more specific for sickle RBC.
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