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Assessment of erythroid dysplasia by “Difference from normal” in routine clinical flow cytometry workup
Author(s) -
Eidenschink Brodersen Lisa,
Menssen Andrew J.,
Wangen Jamie R.,
Stephenson Christine F.,
de Baca Monica E.,
Zehentner Barbara K.,
Wells Denise A.,
Loken Michael R.
Publication year - 2015
Publication title -
cytometry part b: clinical cytometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.646
H-Index - 61
eISSN - 1552-4957
pISSN - 1552-4949
DOI - 10.1002/cyto.b.21199
Subject(s) - myelodysplastic syndromes , dysplasia , myeloid , flow cytometry , pathology , erythropoiesis , biology , phenotype , bone marrow , ficoll , leukemia , immunology , medicine , anemia , genetics , peripheral blood mononuclear cell , gene , in vitro
While multidimensional flow cytometry (MDF) has great utility in diagnostic workups of patients with suspected myelodysplastic syndromes (MDS), only the myeloid lineage has demonstrated reproducible abnormalities from multiple laboratories. With the effects of ammonium chloride (NH 4 Cl) lysis on erythroid progenitors previously described, we applied this protocol to a patient cohort with diagnosed MDS to investigate phenotypic abnormalities that indicate erythroid dysplasia. Method Bone marrow specimens [39 MDS, 9 acute myeloid leukemia (AML), 7 JAK2 V617F positive myeloproliferative neoplasms (MPN), and 5 nutritional deficiencies] were processed by NH 4 Cl lysis and Ficoll preparation and evaluated by MDF using a difference from normal algorithm. Results For the MDS cohort, phenotypic abnormalities on the mature erythroid progenitors were frequent for CD71 and CD36 (36% for each antigen); abnormalities for CD235a (8%) were observed. Among immature erythroid progenitors, abnormal maturation patterns (≤5%), and increased CD105 intensity (9%) were seen. Increased frequency of CD105 bright cells was observed (18%). While antigenic abnormalities correlated between NH 4 Cl lysis and Ficoll preparation, the lysis method demonstrated the most consistent quantitative antigen intensities. Mean erythroid phenotypic abnormalities and prognostic cytogenetic subgroups correlated strongly. Morphologic and erythroid phenotypic abnormalities correlated, as did increasing FCSS and number of erythroid abnormalities, albeit without further increase for AML patients. Discussion These data expand the understanding of erythropoiesis and define immunophenotypic abnormalities that indicate dyserythropoiesis in MDS using a lysis protocol practical for routine implementation in clinical flow cytometric workup. Preliminary studies also indicate strong correlation between phenotypic erythroid dysplasia and poor prognosis, as classified cytogenetically. © 2014 International Clinical Cytometry Society