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
. 2022 Mar 21;14(6):1583.
doi: 10.3390/cancers14061583.

Immunophenotypic Analysis of Acute Megakaryoblastic Leukemia: A EuroFlow Study

Nienke Brouwer  1 , Sergio Matarraz  2   3 , Stefan Nierkens  4 , Mattias Hofmans  5 , Michaela Nováková  6 , Elaine Sobral da Costa  7 , Paula Fernandez  8 , Anne E Bras  1 , Fabiana Vieira de Mello  7 , Ester Mejstrikova  6 , Jan Philippé  9 , Georgiana Emilia Grigore  10 , Carlos E Pedreira  11 , Jacques J M van Dongen  12   13 , Alberto Orfao  2   3 , Vincent H J van der Velden  1 , On Behalf Of The EuroFlow Consortium
Affiliations

Immunophenotypic Analysis of Acute Megakaryoblastic Leukemia: A EuroFlow Study

Nienke Brouwer et al. Cancers (Basel). .

Abstract

Acute megakaryoblastic leukemia (AMKL) is a rare and heterogeneous subtype of acute myeloid leukemia (AML). We evaluated the immunophenotypic profile of 72 AMKL and 114 non-AMKL AML patients using the EuroFlow AML panel. Univariate and multivariate/multidimensional analyses were performed to identify most relevant markers contributing to the diagnosis of AMKL. AMKL patients were subdivided into transient abnormal myelopoiesis (TAM), myeloid leukemia associated with Down syndrome (ML-DS), AML-not otherwise specified with megakaryocytic differentiation (NOS-AMKL), and AMKL-other patients (AML patients with other WHO classification but with flowcytometric features of megakaryocytic differentiation). Flowcytometric analysis showed good discrimination between AMKL and non-AMKL patients based on differential expression of, in particular, CD42a.CD61, CD41, CD42b, HLADR, CD15 and CD13. Combining CD42a.CD61 (positive) and CD13 (negative) resulted in a sensitivity of 71% and a specificity of 99%. Within AMKL patients, TAM and ML-DS patients showed higher frequencies of immature CD34+/CD117+ leukemic cells as compared to NOS-AMKL and AMKL-Other patients. In addition, ML-DS patients showed a significantly higher expression of CD33, CD11b, CD38 and CD7 as compared to the other three subgroups, allowing for good distinction of these patients. Overall, our data show that the EuroFlow AML panel allows for straightforward diagnosis of AMKL and that ML-DS is associated with a unique immunophenotypic profile.

Keywords: AMKL; Down syndrome; EuroFlow; immunophenotyping; transient abnormal myelopoiesis.

PubMed Disclaimer

Conflict of interest statement

J.J.M. van Dongen, J.A. Orfao de Matos Correia e Vale, J.A. Flores-Montero, C.E. Pedreira and V.H.J. van der Velden each report being one of the inventors on the EuroFlow-owned patent PCT/NL2010/050332 (Methods, reagents and kits for flowcytometric immunophenotyping of normal, reactive and malignant leukocytes). The Infinicyt software is based on intellectual property (IP) of some EuroFlow laboratories (University of Salamanca in Spain) and the scientific input of other EuroFlow members. All abovementioned intellectual property and related patents are licensed to Cytognos (Salamanca, ES) and BD Biosciences (San José, CA), which companies pay royalties to the EuroFlow Consortium. These royalties are exclusively used for continuation of the EuroFlow collaboration and sustainability of the EuroFlow consortium. V.H.J.v.d.V. reports a Laboratory Services Agreement with BD Biosciences, Cytognos and Agilent; all related fees are for the Erasmus MC. J.J.M. van Dongen and A. Orfao report an Educational Services Agreement from BD Biosciences (San José, CA) and a Scientific Advisor Agreement with Cytognos; all related fees and honoraria are for the involved university departments at Leiden University Medical Center and University of Salamanca. Georgiana Grigore is an employee of Cytognos. The other authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Figures

Figure 1
Figure 1
Multivariate analysis of non-AMKL (green dots) and AMKL patients (red dots) using the MFI values of all markers present in EuroFlow AML tubes 1–7 (A) or tubes 1–6 (B). Pattern classification was performed using NAPS, and the markers contributing to the pattern classification are shown in the bottom part of the figure. The three AMKL patients not expressing CD42a.CD61, CD41 or CD42b (MFI < 1000) are indicated by arrows.
Figure 2
Figure 2
Differentiation of CD34+/CD117+ AML cells in AMKL (red bars) and non-AMKL patients (green bars). Percentage of positive cells, defined as cells with an MFI > 1000 (mean ± SEM). Differentiation towards B-cell (CD19+), megakaryocytic/erythroid (CD36+/CD64−), erythroid (CD105+), monocytic (CD64+), granulocytic (CD15+) and megakaryocytic (CD42a.CD61+) lineage is shown.
Figure 3
Figure 3
Immunophenotypic profile of CD34+/CD117+, CD34−/CD117+, CD34+/CD117− or CD34−/CD117− subsets of AMKL cells. Expression of markers is depicted as log10 transformed MFI data. Statistical analysis was performed by the Kruskal–Wallis test, followed by the Mann–Whitney test if p < 0.05. The horizontal lines between populations represent statistically significant differences (p < 0.05). The grey zone indicates MFI levels < 1000; markers with such MFI values were considered to be negative.
Figure 4
Figure 4
Univariate analysis of marker expression between the four AMKL subgroups. Data represent the MFI values after log10 transformation. The lines the on top of the figures represent statistically significant differences (p < 0.05) between the two groups. The grey zone indicates MFI levels < 1000; markers with such MFI values were considered to be negative.
Figure 5
Figure 5
Distribution of AMKL cells over the various maturation stages, as defined by CD34 and CD117 expression. The percentage of CD34+/CD117+ leukemic cells was significantly higher in TAM and ML-DS patients as compared to NOS-AMKL and AMKL-Other patients (p < 0.05 by Mann–Whitney test); in contrast, the percentage of CD34−/CD117− leukemic cells was higher in the NOS-AMKL patients (p < 0.05) and AMKL-Other patients (not significant).
Figure 6
Figure 6
Multivariate analysis of marker expression between the four AMKL subgroups. (A) TAM versus ML-DS (contributing markers: CD7 (47%), CD11b (31%) and CD13 (21%)); (B) TAM versus NOS-AMKL (contributing markers: CD117 (28%), CD4 (21%) and CD42b (19%)); (C) TAM versus AMKL-Other (contributing markers: CD117 (61%), CD13 (36%) and CD19 (3%)); (D) ML-DS versus NOS-AMKL (contributing markers: CD7 (68%), CD117 (27%) and CD11b (3%)); (E) ML-DS versus AMKL-Other (contributing markers: CD33 (51%), CD203c (28%) and CD38 (21%)); (F) NOS-AMKL versus AMKL-Other (contributing markers: CD7 (97%), CD38 (2%) nad CD117 (0.4%)). The arrows indicate NOS-AMKL patients with t(1;22).
See this image and copyright information in PMC

References

    1. Arber D.A., Baumann I., Niemeyer C., Brunning R.D., Porwit A. WHO Classification of Tumours Iof Haematopoietic and Lymphoid Tissue. International Agency for Research on Cancer; Lyon, France: 2017. Myeloid proliferations associated with Down Syndrome; pp. 169–171.
    1. Arber D.A., Brunning R.D., Le Beau M.M., Falini B., Vardiman J.W., Porwit A., Thiele J., Foucar K., Dohner H., Bloomfield C.D. WHO Classification of Tumours Iof Haematopoietic and Lymphoid Tissue. International Agency for Research on Cancer; Lyon, France: 2017. Acute myeloid leukemia with recurrent genetic abnormalities; pp. 130–149.
    1. Arber D.A., Brunning R.D., Orazi A., Bain B.J., Porwit A., Le Beau M.M., Greenberg P. WHO Classification of Tumours Iof Haematopoietic and Lymphoid Tissue. International Agency for Research on Cancer; Lyon, France: 2017. Acute myeloid leukemia with myelodysplasia-related changes; pp. 150–152.
    1. Arber D.A., Brunning R.D., Orazi A., Porwit A., Peterson L.C., Thiele J., Le Beau M.M., Hasserjian R.P. WHO Classification of Tumours Iof Haematopoietic and Lymphoid Tissue. International Agency for Research on Cancer; Lyon, France: 2017. Acute myeloid leukemia, NOS; pp. 156–166.
    1. Oki Y., Kantarjian H.M., Zhou X., Cortes J., Faderl S., Verstovsek S., O’Brien S., Koller C., Beran M., Bekele B.N., et al. Adult acute megakaryocytic leukemia: An analysis of 37 patients treated at M.D. Anderson Cancer Center. Blood. 2006;107:880–884. doi: 10.1182/blood-2005-06-2450. - DOI - PubMed