Flow cytometry in myelodysplastic syndrome: analysis of diagnostic utility using maturation pattern-based and quantitative approaches

Abstract

Flow cytometry (FCM) is being increasingly evaluated for the diagnosis of myelodysplastic syndrome (MDS). We employed multiple FCM approaches to assess MDS. Five-color FCM, morphology blind, was done on bone marrow aspirates of 57 suspected MDS and 31 normal controls. Maturation pattern, quantitative FCM for low-grade MDS that awards FCM score, and expression of selected antigens on erythroid cells and CD34(+) blasts were evaluated. FCM results were correlated with clinical and laboratory workup. Patients (n = 57) included proven MDS (n = 14), suspected MDS (n = 13), and non-MDS (n = 30). By pattern-based approach, all proven cases were FCM positive. In suspected MDS, 11 (84.61 %) were positive including morphology-negative cases, and two (15.38 %) were intermediate. In non-MDS cases, 27 of 30 (90 %) were FCM negative, 2 of 30 (6.67 %) intermediate, and 1 of 30 (3.33 %) a hematinic-responsive case, positive. Quantitative parameters that characterized MDS included FCM score of >3, percentage CD34(+) B cells, and expression of CD11b, CD15, and CD56 on myeloblasts. CD71 MFI on CD235a(+) erythroblasts and CD38 MFI on myeloblasts were significantly lower in MDS. The former was present in FCM-intermediate suspected MDS but not FCM-intermediate non-MDS cases. Used in the overall clinical context, both maturation pattern recognition and quantitative approaches, the latter for low-grade MDS, are sensitive methods of diagnosing MDS, including cases negative by morphology and cytogenetics, especially if combined with evaluation of selected antigens, CD71 on CD235a(+) cells and CD38 on CD34(+) cells. The value of FCM in morphology-negative cases needs better definition of specificity through more extensive evaluation of secondary dyspoiesis.

Multicentric study underlining the interest of adding CD5, CD7 and CD56 expression assessment to the flow cytometric Ogata score in myelodysplastic syndromes and myelodysplastic/myeloproliferative neoplasms

“…However, these protocols used a wide diversity of techniques and antibody panels. 9,11,[17][18][19][20][21] With a view to standardizing MFC in MDS, the European LeukemiaNet (ELN) working group on MDS has defined minimal prerequisites for MFC diagnosis of MDS. 21,22 These include markers of myeloid progenitors allowing detection of an abnormal expression of CD45, CD34, CD117, HLA-DR, CD13 or CD33, asynchronous expression of CD11b or CD15 and ectopic expression of CD5, CD7, CD56 or CD19.21,22 Some intracellular markers have also been assessed in MDS, such as myeloperoxidase activity 23 or, more recently, ferritin content 24 or expression of the myeloid nuclear differentiation antigen (MNDA).…”

Flow cytometric detection of dyserythropoiesis: a sensitive and powerful diagnostic tool for myelodysplastic syndromes

“…[5][6][7][8][9] Indeed, several authors have emphasized the specificity and consistency of decreased CD71 expression in MDS patients. 8,10,11 An important caveat in the quantification of erythroid precursors is the modifications induced by red blood cell (RBC) lysis. Elimination of mature non-nucleated erythroid cells is routinely performed in MFC with various reagents.…”

Immunophenotypic Profile of CD34+ Subpopulations and Their Role in the Diagnosis and Prognosis of Patients with De‐Novo, Particularly Low‐Grade Myelodysplastic Syndromes

“…In particular, we investigated (a) CD34+ CD45weak cells which are often analyzed for diagnostic purposes using usually myeloid markers (10)(11)(12)(13)(14)(15)(16)(17)(18)(19)(20) and (b) CD34+ NP, which represent the first maturational steps of granulocytic cells upon attainment of primary granules and which show dysplastic characteristics morphologically and phenotypically in MDS, especially as the disease risk increases (23). In particular, we investigated (a) CD34+ CD45weak cells which are often analyzed for diagnostic purposes using usually myeloid markers (10)(11)(12)(13)(14)(15)(16)(17)(18)(19)(20) and (b) CD34+ NP, which represent the first maturational steps of granulocytic cells upon attainment of primary granules and which show dysplastic characteristics morphologically and phenotypically in MDS, especially as the disease risk increases (23).…”

“…Many studies using MFC have demonstrated abnormalities in the expression of specific antigens such as high CD34 and CD117, as well as abnormal CD45 expression (10)(11)(12)(13)(14)(15)(16). Many studies using MFC have demonstrated abnormalities in the expression of specific antigens such as high CD34 and CD117, as well as abnormal CD45 expression (10)(11)(12)(13)(14)(15)(16).…”

“…MFC has already been used in acute leukemia and lymphoproliferative disorders extensively, but there is not yet consensus regarding MDS. Many studies using MFC have demonstrated abnormalities in the expression of specific antigens such as high CD34 and CD117, as well as abnormal CD45 expression (10)(11)(12)(13)(14)(15)(16). Furthermore, the immunophenotypic characteristics of CD34+ BM cells from MDS patients were analyzed and expansion of neutrophil-lineage CD34+ cells was found in lower risk MDS while expansion of immature CD34+ precursors was observed in higher risk MDS, as well as, in the process of transformation from lower risk MDS with normal karyotype to abnormal karyotype and eventually to higher risk MDS (17)(18)(19)(20).…”