Reticulocyte-gated flow cytometric analysis of red blood cells in paroxysmal nocturnal hemoglobinuria.
Paroxysmal nocturnal hemoglobinuria (PNH) is a hematopoietic stem cell disorder characterized by the deficiency of glycosyl phosphatidylinositol (GPI)-anchored proteins in the affected blood cell membranes. Analysis of blood cells by flow cytometry is useful to identify the affected blood cells with PNH-specific phenotypes. Because PNH-affected red blood cells (RBC) have shortened life-spans in the circulation, ratios of PNH-affected populations analyzed by flow cytometry in whole RBC are lower than those in PNH-affected erythropoiesis. We developed a reticulocyte-gated 2-color flow cytometry of RBC and revealed that the percentages of PNH-affected, CD59--populations in reticulocytes were higher than in whole RBC in patients with PNH. A serial analysis of a patient with PNH who underwent peripheral blood stem cell transplantation confirmed the usefulness of this method to evaluate PNH-affected RBC populations with high sensitivity; ie, the presence of CD59- reticulocytes in the circulation could be a sensitive marker for PNH-affected erythropoiesis.
Sato
Shoko
S
Academic Service Office for Comprehensive Human Sciences, University of T'sukuba, Tsukuba, Japan.
Hasegawa
Yuichi
Y
Nagasawa
Toshiro
T
Ninomiya
Haruhiko
H
D018957:CD59 Antigens / Q000032:analysis; D004906:Erythrocyte Count; D004920:Erythropoiesis; D005434:Flow Cytometry / Q000379:methods*; D006457:Hemoglobinuria, Paroxysmal / Q000097:blood* / Q000175:diagnosis; D006801:Humans; D017701:Reticulocyte Count / Q000295:instrumentation / Q000379:methods; D012156:Reticulocytes / Q000473:pathology*
D018957:CD59 Antigens