Flow cytometric analysis of fetal hemoglobin in erythroid precursors of β-thalassemia

Fetal hemoglobin (HbF), the major hemoglobin species in fetal life, drops to <1% in normal adults. where it is restricted to a few 'F-cells', which may increase in various acquired and genetic conditions, including thalassemia. Using flow cytometry, we studied the percentage of HbF-containing cells and their HbF content in RBC, reticulocytes (retics) and normoblasts (NRBC) present in the peripheral blood of patients with beta-thalassemia. Thiazol orange. a nucleic acid-specific dye, and anti-CD45 antibodies identified the various blood cells and antihuman HbF antibodies quantitated HbF. The results indicated that F-RBC were more numerous in beta-thalassemic (both transfused and nontransfused) patients than in normal donors, but. in most cases, their HbF content was comparable. suggesting that increased HbF in thalassemia is mainly due to higher %F-cells rather than an increased HbF per cell. Among the retics. the %F-cells and their HbF content were highest in immature retics and decreased with maturation to levels of RBC. This may reflect preferential maturation of F-retics into RBC in the circulation. The NRBC population contained the lowest %F-cells. This could be due to preferential maturation of F-NRBC, having more normal phenotype than non-F-NRBC. in the bone marrow into F-retics, while non-F-NRBC enter the circulation.