BACKGROUND: The diagnosis of paroxysmal nocturnal hemoglobinuria (PNH) relies on flow cytometric demonstration of loss of glycosyl-phosphatidyl inositol (GPI)-anchored proteins from red blood cells (RBC) and white blood cells (WBC). High-sensitivity multiparameter assays have been developed to detect loss of GPI-linked structures on PNH neutrophils and monocytes. High-sensitivity assays to detect PNH phenotypes in RBCs have also been developed that rely on the loss of GPI-linked CD59 on CD235a-gated mature RBCs. The latter is used to delineate PNH Type III (total loss of CD59) and PNH Type II RBCs (partial loss of CD59) from normal (Type I) RBCs. However, it is often very difficult to delineate these subsets, especially in patients with large PNH clones who continue to receive RBC transfusions, even while on eculizumab therapy. METHODS: We have added allophycocyanin (APC)-conjugated CD71 to the existing CD235aFITC/CD59PE RBC assay allowing simultaneous delineation and quantification of PNH Type III and Type II immature RBCs (iRBCs). RESULTS: We analyzed 24 medium to large-clone PNH samples (>10% PNH WBC clone size) for PNH Neutrophil, PNH Monocyte, Type III and Type II PNH iRBCs, and where possible, Type III and Type II PNH RBCs. The ability to delineate PNH Type III, Type II, and Type I iRBCs was more objective compared to that in mature RBCs. Additionally, total PNH iRBC clone sizes were very similar to PNH WBC clone sizes. CONCLUSIONS: Addition of CD71 significantly improves the ability to analyze PNH clone sizes in the RBC lineage, regardless of patient hemolytic and/or transfusion status.

• MeSH
• antigeny CD59 metabolismus   MeSH
• buněčná diferenciace MeSH
• CD antigeny krev   fyziologie   MeSH
• diferenciální diagnóza MeSH
• erytrocyty metabolismus   patologie   MeSH
• glykoforin metabolismus   MeSH
• imunofenotypizace přístrojové vybavení   metody   normy   MeSH
• kohortové studie MeSH
• leukocyty patologie   MeSH
• lidé MeSH
• monocyty metabolismus   patologie   MeSH
• neutrofily metabolismus   patologie   MeSH
• paroxysmální hemoglobinurie krev   klasifikace   diagnóza   patologie   MeSH
• počet leukocytů přístrojové vybavení   metody   MeSH
• průtoková cytometrie přístrojové vybavení   metody   normy   MeSH
• receptory transferinu krev   fyziologie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH