Premium
Screening bone marrow samples for abnormal lymphoid populations and myelodysplasia‐related features with one 10‐color 14‐antibody screening tube
Author(s) -
Rajab Amr,
Porwit Anna
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.21233
Subject(s) - medicine , myeloid , cd34 , bone marrow , cd5 , myelodysplastic syndromes , cd3 , cd19 , lymphocyte , cd33 , cd20 , antibody , gastroenterology , immunology , cd8 , pathology , antigen , biology , stem cell , genetics
Background We have designed one‐tube 14‐antibody 10‐color flow cytometry (FCM) panel that would provide maximum information on lymphoid and myeloid cell subsets in bone marrow aspirates (BMA) from patients with cytopenia(s). Samples and Methods BMA from 8 normal donors, from 286 non‐malignant hospital controls, 92 myelodysplastic syndromes (MDS), 47 myeloproliferative neoplasms (MPN), and from 14 MDS/MPN patients were investigated. One tube 14‐monoclonal antibody (MAb) 10‐fluorochrome panel included: kappa+CD4 FITC, Lambda+CD8 PE, CD3 + CD14 ECD, CD34 APC, CD20+CD56 PC7, CD10 APC‐A750, CD19 APC‐A700, CD33 PC5.5, CD5 PB, and CD45 KO. Kappa/lambda expression was evaluated separately in CD19+, CD10+ and CD5+ B‐cells. CD4+CD3+, CD8+CD3+, CD5+CD3+ T‐lymphocyte subsets were enumerated. Blasts were evaluated using CD45/SSC and CD34 gating. The FCM score for MDS (sc. Ogata score) included CD34+ myeloblast and B‐progenitor cluster size, myeloblast/lymphocyte CD45 expression, and granulocyte/lymphocyte SSC ratio. Results Abnormal lymphoid populations or increased plasma cells were found in 18 patients (4%). A 43/92 BMA from MDS and 7/14 from MDS/MPN patients had score >2. Score >2 had 92.5% positive predictive value for MDS/MDS‐MPN diagnosis. Negative predictive value for MDS/MDS‐MPN was 83% for scores under 3 and 88% for scores under 2. All but two of normal/hospital control samples had FCM score <3 (99%). Differences in scores between MDS & MDS/MPN and the control groups were statistically significant ( P < 0.0001). Conclusions Our one‐tube FCM panel can be easily applied to screening for aberrant lymphoid populations and myelodysplasia‐related features. MDS scores >2 are highly indicative of MDS or MDS‐MPN. © 2015 International Clinical Cytometry Society