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. 2019 Mar;96(2):134-142.
doi: 10.1002/cyto.b.21745. Epub 2018 Nov 18.

CD123 expression levels in 846 acute leukemia patients based on standardized immunophenotyping

Anne E Bras  1 , Valerie de Haas  2 , Arthur van Stigt  3 , Mojca Jongen-Lavrencic  4 , H Berna Beverloo  5 , Jeroen G Te Marvelde  1 , C Michel Zwaan  6   7 , Jacques J M van Dongen  1 , Jeanette H W Leusen  3 , Vincent H J van der Velden  1
Affiliations

CD123 expression levels in 846 acute leukemia patients based on standardized immunophenotyping

Anne E Bras et al. Cytometry B Clin Cytom. 2019 Mar.

Abstract

Background: While it is known that CD123 is normally strongly expressed on plasmacytoid dendritic cells and completely absent on nucleated red blood cells, detailed information regarding CD123 expression in acute leukemia is scarce and, if available, hard to compare due to different methodologies.

Methods: CD123 expression was evaluated using standardized EuroFlow immunophenotyping in 139 pediatric AML, 316 adult AML, 193 pediatric BCP-ALL, 69 adult BCP-ALL, 101 pediatric T-ALL, and 28 adult T-ALL patients. Paired diagnosis-relapse samples were available for 57 AML and 19 BCP-ALL patients. Leukemic stem cell (LSC) data was available for 32 pediatric AML patients. CD123 expression was evaluated based on mean fluorescence intensity, median fluorescence intensity, and percentage CD123 positive cells.

Results: EuroFlow panels were stable over time and between laboratories. CD123 was expressed in the majority of AML and BCP-ALL patients, but absent in most T-ALL patients. Within AML, CD123 expression was lower in erythroid/megakaryocytic leukemia, higher in NPM1 mutated and FLT3-ITD mutated leukemia, and comparable between LSC and leukemic blasts. Within BCP-ALL, CD123 expression was higher in patients with (high) hyperdiploid karyotypes and the BCR-ABL fusion gene. Interestingly, CD123 expression was increased in BCP-ALL relapses while highly variable in AML relapses (compared to CD123 expression at diagnosis).

Conclusions: Authors evaluated CD123 expression in a large cohort of acute leukemia patients, based on standardized and reproducible methodology. Our results may facilitate stratification of patients most likely to respond to CD123 targeted therapies and serve as reference for CD123 expression (in health and disease). © 2018 The Authors. Cytometry Part B: Clinical Cytometry published by Wiley Periodicals, Inc. on behalf of International Clinical Cytometry Society.

Keywords: CD123; acute leukemia; standardized immunophenotyping; targeted therapy.

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Conflict of interest statement

None were disclosed by the authors.

Figures

Figure 1
Figure 1
Assay stability over time (2011–2016) and across laboratories (EMC and DCOG). Evaluation based on normal NRBC and normal PDC (as negative and positive control, respectively). Visualization per individual population, where the horizontal gray bars visualize the 96% interval and the vertical black bars visualize the median CD123 expression (CD123‐MFI). Solid vertical line indicates the highest 98 percentile found among all NRBC populations, which was used as positivity cut‐off during CD123‐PPC analysis. For each subgroup, the Wilcoxon signed‐rank test result is shown, comparing the subgroup against the remainder of populations (Wp denotes the probability value after Bonferroni correction). One‐way ANOVA did not reveal any significant differences among the NRBC and PDC subgroups as well (both tests P > 0.05). The Bartlett test, Fligner‐Killeen test, and Brown–Forsythe tests, comparing all PDC subgroups at once, did not reveal any significant differences as well (P = 0.154, P = 0.065, P = 0.075, respectively). In conclusion, evaluated subgroups have comparable distributions and variances, confirming assay stability over time and across centers.
Figure 2
Figure 2
CD123 median fluorescence intensity (CD123‐MFI) for normal mature leukocyte subsets and acute leukemia. Curved gray lines visualize Gaussian kernel density estimation. Vertical gray bars visualize performed Wilcoxon signed‐rank tests (P‐value after Bonferroni multiple testing correction was smaller than 0.001 unless otherwise specified). (A) CD123‐MFI for AML, BCP‐ALL, and T‐ALL patients and various normal populations. Vertical dotted line represents the highest CD123‐MFI among evaluated NRBC populations. (B) CD123‐MFI levels for selected AML subgroups. (C) CD123‐MFI levels for selected BCP‐ALL subgroups.
Figure 3
Figure 3
Percentage CD123 positive cells (CD123‐PPC) for AML, BCP‐ALL, and T‐ALL patients. Percentage CD123 positive cells based on 739 intensity cut‐off. Black lines visualize pediatric patients and gray lines visualize adult patients. Gray area visualizes the difference between pediatric and adult patients (reported P‐values based on a Mann–Whitney U tests). Other immunophenotyping studies typically state “at least … patients have at least … percent positive cells.” As different studies use different percentages, we have chosen for this visualization, which allows lookup of any combination. For example, approximately 65% of pediatric AML patients have at least 60% CD123 positive cells based on the 739 positivity cut‐off. (A) CD123‐PPC for AML, BCP‐ALL and T‐ALL patients. (B) CD123‐PPC for selected AML subgroups. (C) CD123‐PPC for selected BCP‐ALL subgroups.
Figure 4
Figure 4
Paired evaluations (diagnosis versus relapse and leukemic blasts vs. leukemic stem cells). Initial diagnosis (DX) versus relapse (RX) for BCP‐ALL patients (A) and AML patients (B). Leukemic blasts versus LSCs in AML patients (C). Black bars indicate increased CD123‐MFI while gray bars indicate decreased CD123‐MFI (right vs. left side). Lower panels visualize the delta CD123 expression (after default logicle transformation), thereby clearly visualizing the magnitude and direction of changes. Reported P‐values are based on a paired Mann–Whitney U test.
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References

    1. Munoz L, Nomdedeu JF, Lopez O, Carnicer MJ, Bellido M, Aventin A, Brunet S, Sierra J. Interleukin‐3 receptor alpha chain (CD123) is widely expressed in hematologic malignancies. Haematologica. 2001;86:1261–1269. - PubMed
    1. Testa U, Pelosi E, Frankel A. CD 123 is a membrane biomarker and a therapeutic target in hematologic malignancies. Biomark Res. 2014;2:4. - PMC - PubMed
    1. Buckley SA, Walter RB. Update on antigen‐specific immunotherapy of acute myeloid leukemia. Curr Hematol Malig Rep. 2015;10:65–75. - PubMed
    1. Garfin PM, Feldman EJ. Antibody‐based treatment of acute myeloid leukemia. Curr Hematol Malig Rep. 2016;11:545–552. - PubMed
    1. Nijhof IS, Casneuf T, van Velzen J, van Kessel B, Axel AE, Syed K, Groen RW, van Duin M, Sonneveld P, Minnema MC, et al. CD38 expression and complement inhibitors affect response and resistance to daratumumab therapy in myeloma. Blood. 2016;128:959–970. - PubMed

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