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Screening bone marrow samples for abnormal lymphoid populations and myelodysplasia-related features with one 10-color 14-antibody screening tube

Rajab, Amr and Porwit, Anna LU (2015) In Cytometry Part B - Clinical Cytometry 88(4). p.253-260
Abstract

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+,... (More)

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.

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author
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publishing date
type
Contribution to journal
publication status
published
keywords
bone marrow, flow cytometry, lymphoma, myelodysplastic syndrome
in
Cytometry Part B - Clinical Cytometry
volume
88
issue
4
pages
8 pages
publisher
John Wiley & Sons Inc.
external identifiers
  • scopus:84932200033
  • pmid:25664445
ISSN
1552-4949
DOI
10.1002/cyto.b.21233
language
English
LU publication?
no
id
4a901211-6f5f-4407-a3e1-55c8ec6403c1
date added to LUP
2019-05-22 09:44:27
date last changed
2024-04-16 07:21:18
@article{4a901211-6f5f-4407-a3e1-55c8ec6403c1,
  abstract     = {{<p>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 &gt;2. Score &gt;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 &lt;3 (99%). Differences in scores between MDS &amp; MDS/MPN and the control groups were statistically significant (P-&lt;-0.0001). Conclusions Our one-tube FCM panel can be easily applied to screening for aberrant lymphoid populations and myelodysplasia-related features. MDS scores &gt;2 are highly indicative of MDS or MDS-MPN.</p>}},
  author       = {{Rajab, Amr and Porwit, Anna}},
  issn         = {{1552-4949}},
  keywords     = {{bone marrow; flow cytometry; lymphoma; myelodysplastic syndrome}},
  language     = {{eng}},
  month        = {{07}},
  number       = {{4}},
  pages        = {{253--260}},
  publisher    = {{John Wiley & Sons Inc.}},
  series       = {{Cytometry Part B - Clinical Cytometry}},
  title        = {{Screening bone marrow samples for abnormal lymphoid populations and myelodysplasia-related features with one 10-color 14-antibody screening tube}},
  url          = {{http://dx.doi.org/10.1002/cyto.b.21233}},
  doi          = {{10.1002/cyto.b.21233}},
  volume       = {{88}},
  year         = {{2015}},
}