title: The distribution of PIG-A gene abnormalities in paroxysmal nocturnal hemoglobinuria granulocytes and cultured erythroblasts.
abstract: Paroxysmal nocturnal hemoglobinuria (PNH) is an acquired hemolytic anemia that is characterized by a deficiency of glycosylphosphatidylinositol-anchored membrane proteins due to phosphatidylinositol glycan-class A (PIG-A) gene abnormalities in various lineages of peripheral blood cells and hematopoietic precursors. The purpose of our study was to clarify the distribution of PIG-A gene abnormalities among various cell lineages during differentiation and maturation in PNH patients. The expression of CD16b or CD59 in peripheral blood granulocytes or cultured erythroblasts from three Japanese PNH patients was analyzed using flow cytometry. PIG-A gene abnormalities in both cell types, including glycophorin A(+) bone marrow erythroblasts, were examined using nucleotide sequence analysis. The expression study of PIG-A genes from each patient was also performed using JY-5 cells.Flow cytometry revealed that the erythroblasts consisted of negative, intermediate, and positive populations in Cases 1 and 3 and negative and intermediate populations in Case 2. The granulocytes consisted of negative and positive populations in all three cases. DNA sequence analysis indicated that all the PNH cases had two or three types of PIG-A gene abnormalities, and that a predominant clone with an abnormal PIG-A gene was different in granulocytes and erythroblasts from Cases 2 and 3. Expression studies showed that all the mutations from the patients were responsible for the null phenotype.PIG-A gene abnormalities result in deficiencies of glycosylphosphatidylinositol-anchored proteins in PNH erythroblasts and granulocytes. The distribution of predominant PNH clones with PIG-A gene abnormalities is often heterogeneous between the cell types, suggesting that a clonal selection of PIG-A gene abnormalities occurs independently among various cell lineages during differentiation and maturation.
authors: First Department of Internal Medicine, Fukushima Medical University, 1 Hikariga-oka, Fukushima 960-1295, Japan.
authors: H
authors: H
authors: Noji
authors: T
authors: T
authors: Shichishima
authors: Y
authors: Y
authors: Saitoh
authors: T
authors: T
authors: Kai
authors: T
authors: T
authors: Yamamoto
authors: K
authors: K
authors: Ogawa
authors: M
authors: M
authors: Okamoto
authors: K
authors: K
authors: Ikeda
authors: Y
authors: Y
authors: Maruyama
mesh_terms: D000328:Adult; D000368:Aged; D018958:CD55 Antigens / Q000032:analysis; D018957:CD59 Antigens / Q000032:analysis; D002454:Cell Differentiation; D002478:Cells, Cultured; D004247:DNA / Q000097:blood / Q000737:chemistry; D004900:Erythroblasts / Q000737:chemistry* / Q000276:immunology; D004912:Erythrocytes / Q000737:chemistry; D005260:Female; D005434:Flow Cytometry; D017261:Glycosylphosphatidylinositols / Q000097:blood / Q000172:deficiency; D006098:Granulocytes / Q000737:chemistry* / Q000276:immunology; D006457:Hemoglobinuria, Paroxysmal / Q000097:blood / Q000235:genetics*; D006801:Humans; D008297:Male; D008565:Membrane Proteins / Q000235:genetics*; D009154:Mutation; D012333:RNA, Messenger / Q000097:blood / Q000737:chemistry; D017452:Receptors, IgG / Q000032:analysis; D017422:Sequence Analysis, DNA; D017423:Sequence Analysis, RNA
chemical_list: D018958:CD55 Antigens; D018957:CD59 Antigens; D017261:Glycosylphosphatidylinositols; D008565:Membrane Proteins; D012333:RNA, Messenger; D017452:Receptors, IgG; C080086:phosphatidylinositol glycan-class A protein; D004247:DNA