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Analysis of erythroid maturation in the nonlysed bone marrow with help of radar plots facilitates detection of flow cytometric aberrations in myelodysplastic syndromes

Violidaki, Despoina LU orcid ; Axler, Olof LU ; Jafari, Katayoon ; Bild, Filippa LU ; Nilsson, Lars LU ; Mazur, Joanna ; Ehinger, Mats LU and Porwit, Anna LU (2020) In Cytometry Part B - Clinical Cytometry 98(5). p.399-411
Abstract

Background: Accumulating data support the role of flow cytometry (FCM) in diagnostic work-up of myelodysplastic syndromes (MDS). Changes in erythropoiesis are less documented than in granulopoiesis. However, most studies were performed on bone marrow samples (BMSs) after red blood cell lysis. We have established a FCM protocol for erythropoiesis, following a no-lysis approach and live gate acquisition of nucleated cells using DNA dye DRAQ5. Methods: The ERY tube consisted of CD36, CD71, CD105, CD117, CD13, and CD45. Comparison with cytomorphological differential counts was carried out in a learning cohort of 80 BMS. To detect aberrations, we analyzed 208 BMS from 135 patients and five normal donors, divided into three cohorts: MDS (n =... (More)

Background: Accumulating data support the role of flow cytometry (FCM) in diagnostic work-up of myelodysplastic syndromes (MDS). Changes in erythropoiesis are less documented than in granulopoiesis. However, most studies were performed on bone marrow samples (BMSs) after red blood cell lysis. We have established a FCM protocol for erythropoiesis, following a no-lysis approach and live gate acquisition of nucleated cells using DNA dye DRAQ5. Methods: The ERY tube consisted of CD36, CD71, CD105, CD117, CD13, and CD45. Comparison with cytomorphological differential counts was carried out in a learning cohort of 80 BMS. To detect aberrations, we analyzed 208 BMS from 135 patients and five normal donors, divided into three cohorts: MDS (n = 68), nonclonal cytopenia (n = 43), and normal controls (n = 29). Radar plot (RP) was created for an overview of normal and aberrant patterns. Results: The proportion of erythropoiesis in the ERY tube showed better agreement with the cytomorphology, compared to FCM panels on lysed BMS. We confirmed that aberrations in coefficient of variation (CV) of CD36 fluorescence intensity (p <.001), mean fluorescence intensity of CD36 (p =.012), and CV of CD105 (p <.001) can distinguish between MDS and nonclonal cytopenia. RP facilitated evaluation of erythropoietic maturation patterns and aberrant patterns were identified in 85% of MDS patients. Conclusion: This study provides evidence that a no-lysis approach and RP analysis allow a more reliable evaluation of erythropoiesis and erythroid dysplasia, supporting the integration of FCM erythroid panels in the standard work-up of MDS.

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author
; ; ; ; ; ; and
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
erythroid dysplasia, erythropoiesis, flow cytometry, lysis, myelodysplastic syndrome
in
Cytometry Part B - Clinical Cytometry
volume
98
issue
5
pages
13 pages
publisher
John Wiley & Sons Inc.
external identifiers
  • pmid:32543774
  • scopus:85086447266
ISSN
1552-4949
DOI
10.1002/cyto.b.21931
language
English
LU publication?
yes
id
7cd31a5e-5162-4e61-89d3-5ea86212b58c
date added to LUP
2020-07-06 14:32:54
date last changed
2024-10-03 04:33:36
@article{7cd31a5e-5162-4e61-89d3-5ea86212b58c,
  abstract     = {{<p>Background: Accumulating data support the role of flow cytometry (FCM) in diagnostic work-up of myelodysplastic syndromes (MDS). Changes in erythropoiesis are less documented than in granulopoiesis. However, most studies were performed on bone marrow samples (BMSs) after red blood cell lysis. We have established a FCM protocol for erythropoiesis, following a no-lysis approach and live gate acquisition of nucleated cells using DNA dye DRAQ5. Methods: The ERY tube consisted of CD36, CD71, CD105, CD117, CD13, and CD45. Comparison with cytomorphological differential counts was carried out in a learning cohort of 80 BMS. To detect aberrations, we analyzed 208 BMS from 135 patients and five normal donors, divided into three cohorts: MDS (n = 68), nonclonal cytopenia (n = 43), and normal controls (n = 29). Radar plot (RP) was created for an overview of normal and aberrant patterns. Results: The proportion of erythropoiesis in the ERY tube showed better agreement with the cytomorphology, compared to FCM panels on lysed BMS. We confirmed that aberrations in coefficient of variation (CV) of CD36 fluorescence intensity (p &lt;.001), mean fluorescence intensity of CD36 (p =.012), and CV of CD105 (p &lt;.001) can distinguish between MDS and nonclonal cytopenia. RP facilitated evaluation of erythropoietic maturation patterns and aberrant patterns were identified in 85% of MDS patients. Conclusion: This study provides evidence that a no-lysis approach and RP analysis allow a more reliable evaluation of erythropoiesis and erythroid dysplasia, supporting the integration of FCM erythroid panels in the standard work-up of MDS.</p>}},
  author       = {{Violidaki, Despoina and Axler, Olof and Jafari, Katayoon and Bild, Filippa and Nilsson, Lars and Mazur, Joanna and Ehinger, Mats and Porwit, Anna}},
  issn         = {{1552-4949}},
  keywords     = {{erythroid dysplasia; erythropoiesis; flow cytometry; lysis; myelodysplastic syndrome}},
  language     = {{eng}},
  number       = {{5}},
  pages        = {{399--411}},
  publisher    = {{John Wiley & Sons Inc.}},
  series       = {{Cytometry Part B - Clinical Cytometry}},
  title        = {{Analysis of erythroid maturation in the nonlysed bone marrow with help of radar plots facilitates detection of flow cytometric aberrations in myelodysplastic syndromes}},
  url          = {{http://dx.doi.org/10.1002/cyto.b.21931}},
  doi          = {{10.1002/cyto.b.21931}},
  volume       = {{98}},
  year         = {{2020}},
}