Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2023 Mar;104(2):162-172.
doi: 10.1002/cyto.b.22038. Epub 2021 Nov 22.

Dysplasia and PNH-type cells in bone marrow aspirates of myelodysplastic syndromes

Theresia M Westers  1 , Canan Alhan  1 , Heleen A Visser-Wisselaar  2 , Dana A Chitu  2 , Arjan A van de Loosdrecht  1
Affiliations
Free article

Dysplasia and PNH-type cells in bone marrow aspirates of myelodysplastic syndromes

Theresia M Westers et al. Cytometry B Clin Cytom. 2023 Mar.
Free article

Abstract

Background: Flow cytometry is increasingly applied in cytopenic patients suspected for myelodysplastic syndromes (MDS). Analysis includes evaluation of antigen expression patterns in granulocytes of which, for example, partial lack of CD16 may indicate dysplasia, but presence of paroxysmal nocturnal hemoglobinuria (PNH)-type cells should be considered. However, diagnostic bone marrow (BM) samples hamper PNH analysis because immature stages in the granulo-/monocytic compartment lack expression of certain glycophosphatidyl-inositol-anchored proteins. In this prospective study, we evaluated the presence of PNH-type cells in BM next to aberrancies from routine MDS immunophenotyping.

Methods: We combined antibodies defining maturation trajectories with FLAER. Validation of the designed method against routine PNH analysis and parallel analysis of BM and blood samples revealed similar results (granulocytes: Wilcoxon p = 0.25 and p = 0.82, respectively). We analyzed BM samples from 134 MDS, 17 chronic myelomonocytic leukemia, 15 aplastic anemia (AA), 1 PNH, 51 non-clonal cytopenic controls, and 12 normal controls.

Results: Most AA/PNH-BM samples showed clear PNH clones: median 1.1% (0%-35%); CD16 loss on mature neutrophils paralleled PNH-clone sizes. In MDS-BM, only 3.7% of cases showed ≥0.1% PNH-type cells, whereas partial CD16 loss was more frequent and abundant.

Conclusions: Our findings confirm that dysplastic features in MDS-BM may point to presence of PNH-type cells, though only few cases displayed FLAER-negative cells. We showed that identification of these cells in the granulocyte compartment of BM specimen is feasible, but-according to international guidelines-results need to be confirmed in peripheral blood.

Keywords: MDS; PNH-type cells; bone marrow; dysplasia; flow cytometry.

PubMed Disclaimer

References

REFERENCES

    1. Borowitz, M. J., Craig, F. E., Digiuseppe, J. A., Illingworth, A. J., Rosse, W., Sutherland, D. R., Wittwer, C. T., & Richards, S. J. (2010). Guidelines for the diagnosis and monitoring of paroxysmal nocturnal hemoglobinuria and related disorders by flow cytometry. Cytometry. Part B, Clinical Cytometry, 78, 211-230.
    1. Brando, B., Gatti, A., & Preijers, F. (2019). Flow cytometric diagnosis of paroxysmal nocturnal hemoglobinuria: Pearls and pitfalls-A critical review article. EJIFCC, 30, 355-370.
    1. Cremers, E. M. P., Westers, T. M., Alhan, C., Cali, C., Visser-Wisselaar, H. A., Chitu, D. A., van der Velden, V. H., te Marvelde, J. G., Klein, S. K., Muus, P., Vellenga, E., de Gree, G. E., Legdeur, M. C., Wijermans, P. W., Stevens-Kroef, M. J., Silva-Coelho, P. D., Jansen, J. H., Ossenkoppele, G. J., & van de Loosdrecht, A. A. (2017). Implementation of erythroid lineage analysis by flow cytometry in diagnostic models for myelodysplastic syndromes. Haematologica, 102, 320-326.
    1. de Haas, M., Kleijer, M., van Zwieten, R., Roos, D., & von Dem Borne, A. E. (1995). Neutrophil Fc gamma RIIIb deficiency, nature, and clinical consequences: A study of 21 individuals from 14 families. Blood, 86, 2403-2413.
    1. Dezern, A. E., & Borowitz, M. J. (2018). ICCS/ESCCA consensus guidelines to detect GPI-deficient cells in paroxysmal nocturnal hemoglobinuria (PNH) and related disorders part 1-Clinical utility. Cytometry Part B, Clinical Cytometry, 94, 16-22.

Publication types

MeSH terms

LinkOut - more resources