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  • Title: Bone Marrow as a Source of Cells for Paroxysmal Nocturnal Hemoglobinuria Detection.
    Author: Dulau-Florea AE, Young NS, Maric I, Calvo KR, Dunbar CE, Townsley DM, Winkler T, Monreal M, Jiang C, Jordan EK, Braylan RC.
    Journal: Am J Clin Pathol; 2018 Jul 31; 150(3):273-282. PubMed ID: 29982419.
    Abstract:
    OBJECTIVES: To determine fluorescently labeled aerolysin (FLAER) binding and glycophosphatidylinositol-anchored protein expression in bone marrow (BM) cells of healthy volunteers and patients with paroxysmal nocturnal hemoglobinuria (PNH) detected in peripheral blood (PB); compare PNH clone size in BM and PB; and detect PNH in BM by commonly used antibodies. METHODS: Flow cytometry analysis of FLAER binding to leukocytes and expression of CD55/CD59 in erythrocytes. Analysis of CD16 in neutrophils and CD14 in monocytes in BM. RESULTS: FLAER binds to all normal BM leukocytes, and binding increases with cell maturation. In PNH, lymphocytic clones are consistently smaller than clones of other BM cells. PNH clones are detectable in mature BM leukocytes with high specificity and sensitivity using common antibodies. CONCLUSIONS: PNH clone sizes measured in mature BM leukocytes and in PB are comparable, making BM suitable for PNH assessment. We further demonstrate that commonly used reagents (not FLAER or CD55/CD59) can reliably identify abnormalities of BM neutrophils and monocytes consistent with PNH cells.
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