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Blood, 1 December 2004, Vol. 104, No. 12, pp. 3782-3788.
Prepublished online as a Blood First Edition Paper on August 10, 2004; DOI 10.1182/blood-2004-02-0645.
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Submitted February 20, 2004
Accepted June 24, 2004
Evidence for Transfer of Glycosylphosphatidylinositol-Anchored Proteins to Deficient Cells After Erythrocyte Transfusion in Paroxysmal Nocturnal Hemoglobinuria
Elaine M Sloand*, Lori Mainwaring, Keyvan Keyvanfar, Jichun Chen, Jaroslaw Maciejewski, Harvey G Klein, and Neal S Young
Hematology Branch, National Heart, Lung, and Blood Institute, Bethesda, MD, USA
Department of Transfusion Medicine, Clinical Center, National Institutes of Health, Bethesda, MD, USA
* Corresponding author; email: sloande{at}nih.gov.
In paroxysmal nocturnal hemoglobinuria (PNH), an acquired mutation of the PIG-A gene results in the absence of glycosylphosphoinositol (GPI)-anchored cell surface membrane proteins in affected hematopoietic cells. Absence of GPI-anchored proteins on erythrocytes is responsible for their increased sensitivity to complement-mediated lysis, resulting in hemolytic anemia. Cell-to-cell transfer of CD55 and CD59, two GPI-anchored proteins, by red cell microvesicles has been demonstrated in vitro, with retention of their function. Since red cell units stored for transfusion contain many erythrocyte microvesicles, transfused blood could potentially serve as a source of CD55 and CD59. We examined whether GPI-anchored proteins could be transferred in vivo to deficient cells following transfusion of PNH patients. Our subjects were six patients with severe hemolytic PNH who required frequent transfusions; their blood group was A1. Each was transfused with three units of compatible, washed group O blood. Patient group A1 cells were distinguished from the transfused group O cells by staining with a fluorescein isothiocyanate (FITC)-conjugated lectin, Dolichos biflorus, which specifically binds to group A1 erythrocytes. Cells were labeled with phycoerythrin (PE)-conjugates of monoclonal antibodies to CD55 or CD59 to detect transfer of these proteins using flow cytometry. Increased surface CD59 was measured on recipient red cells and granulocytes one, three, and seven days following transfusion in all six patients. Our data suggest a potential therapeutic role for GPI-anchored protein transfer for severe PNH.
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