The potential of proliferative and apoptotic parameters in clinical flow cytometry of myeloid malignancies

Abstract Standardization of the detection and quantification of leukocyte differentiation markers by the EuroFlow Consortium has led to a major step forward in the integration of flow cytometry into classification of leukemia and lymphoma. In our opinion, this now enables introduction of markers for more dynamic parameters, such as proliferative and (anti)apoptotic markers, which have proven their value in the field of histopathology in the diagnostic process of solid tumors and lymphoma. Although use of proliferative and (anti)apoptotic markers as objective parameters in the diagnostic process of myeloid malignancies was studied in the past decades, this did not result in the incorporation of these biomarkers into clinical diagnosis. This review addresses the potential of these markers for implementation in the current, state-of-the-art multiparameter analysis of myeloid malignancies. The reviewed studies clearly recognize the importance of proliferation and apoptotic mechanisms in the pathogenesis of bone marrow (BM) malignancies. The literature is, however, contradictory on the role of these processes in myelodysplastic syndrome (MDS), MDS/myeloproliferative neoplasms, and acute myeloid leukemia. Furthermore, several studies underline the need for the analysis of the proliferative and apoptotic rates in subsets of hematopoietic BM cell lineages and argue that these results can have diagnostic and prognostic value in patients with myeloid malignancies. Recent developments in multiparameter flow cytometry now allow quantification of proliferative and (anti)apoptotic indicators in myeloid cells during their different maturation stages of separate hematopoietic cell lineages. This will lead to a better understanding of the biology and pathogenesis of these malignancies.

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Multiparameter flow cytometry reveals myelodysplasia-related aberrant antigen expression in myelodysplastic/myeloproliferative neoplasms

Cytometry Part B Clinical Cytometry ◽

10.1002/cyto.b.21068 ◽

2013 ◽

Vol 84B (3) ◽

pp. 194-197 ◽

Cited By ~ 10

Author(s):

Wolfgang Kern ◽

Ulrike Bacher ◽

Susanne Schnittger ◽

Tamara Alpermann ◽

Claudia Haferlach ◽

...

Keyword(s):

Flow Cytometry ◽

Myeloproliferative Neoplasms ◽

Antigen Expression ◽

Multiparameter Flow Cytometry

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Detection of Dysplastic Features by Multiparameter Flow Cytometry in Myelodysplastic Syndromes in Relation to Cytomorphology and Cytogenetics.

Blood ◽

10.1182/blood.v110.11.2461.2461 ◽

2007 ◽

Vol 110 (11) ◽

pp. 2461-2461

Author(s):

Wolfgang Kern ◽

Claudia Haferlach ◽

Susanne Schnittger ◽

Torsten Haferlach

Keyword(s):

Flow Cytometry ◽

Myelodysplastic Syndromes ◽

Erythroid Cell ◽

Multiparameter Flow Cytometry ◽

Cell Populations ◽

Erythroid Cells ◽

Monocytic Cell ◽

Aberrant Expression ◽

Cell Lineages ◽

Cd16 Expression

Abstract Dysplastic features can be detected in different cell lineages in myelodysplastic syndromes (MDS) by multiparameter flow cytometry (MFC). The aim of the present study has been the assessment of the flow cytometric detection of dysplastic features previously published to occur in MDS in relation to findings in cytomorphology (CM) and cytogenetics (CG). We analyzed 307 bone marrow samples from patients with suspected (n=130) or proven (n=177) MDS by MFC, CM, and CG in parallel. Blast counts as determined by CM and MFC, respectively, ranged from 0% to 21% (median, 3.5%) and from 0% to 23% (median, 3%; r=0.271, p<0.0001). The median number of aberrant features detected by MFC were 0 for blasts (range, 0 to 4), 2 for granulocytes (0 to 5), 1 for monocytes (0 to 5), and 0 for erythroid cells (0 to 2); median total number=3, range 0–11. The most frequent dysplastic features observed in the blast populations included aberrant coexpression of CD11b (13.7%), CD15 (10.4%) and CD64 (10.4%). The most frequent dysplastic features observed in the granulocytic cell populations included reduced side-scatter signal corresponding to hypogranulation (67.1%), aberrant coexpression of CD56 (32.9%), aberrant pattern of CD13/CD16 expression (31.6%), aberrant pattern of CD11b/CD16 expression (24.1%), and reduced expression of CD33 (13.4%). The most frequent dysplastic features observed in the monocytic cell populations included aberrant coexpression of CD56 (42.7%) and of CD16 (18.9%). The most frequent dysplastic features observed in the erythroid cell populations included a lack of CD71 expression (15.0%) and an aberrantly homogeneous expression of CD71 (9.1%). As compared to cases with no indication of MDS by CM (=non-MDS) cases with MDS according to CM were significantly associated with a reduced side-scatter signal in granulocytes (ratio granulocytes:lymphocytes 6.53±1.27 vs. 7.44±1.17, p<0.0001) as well as a higher number of dysplastic features in granulocytes (1.98±1.09 vs. 1.00±1.31, p<0.0001), monocytes (0.81±0.84 vs. 0.35±0.63, p<0.0001), and erythroid cells (0.33±0.47 vs. 0.20±0.40, p=0.061). Particularly, an aberrant expression of CD56 in monocytes occurred more frequently in 33 cases with CMML as compared to non-MDS cases (84.8% vs. 15.7%, p<0.0001). In cases with possible MDS according to CM the differences to non-MDS cases were less pronounced (reduced side-scatter signal in granulocytes 7.77±1.47 vs. 7.44±1.17, n.s., dysplastic features in granulocytes 1.90±1.21 vs. 1.00±1.31, p=0.003, monocytes 0.50±0.63 vs. 0.35±0.63, n.s., and erythroid cells 0.30±0.47 vs. 0.20±0.40, n.s.). In cases with aberrant cytogenetics (n=80, excluding those with –Y as sole aberration) dysplastic features by MFC occurred more frequently as compared to cases with normal karyotypes (reduced side-scatter signal in granulocytes 6.35±1.18 vs. 7.09±1.38, p<0.0001, dysplastic features in granulocytes 2.05±0.95 vs. 1.68±1.31, p=0.021, monocytes 0.80±0.89 vs. 0.71±0.81, n.s., and erythroid cells 0.38±0.49 vs. 0.31±0.48, n.s.). A total of more than two dysplastic features in blasts, granulocytes, and monocytes and/or a blast count >5% occurred in 68.4%, 63.3%, and 35.3% of cases with MDS, possible MDS, and non-MDS according to CM (p=0.0005). Interestingly, in some cases aberrant antigen expression has been observed in cell lineages not rated dysplastic by CM. This evaluation suggests that MFC may be used to identify dysplastic features in patients with suspected MDS. Sensitivity and specificity varies between MDS subtypes and should be clearly defined in future studies.

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The importance of evaluating specific myeloid malignancies in epidemiological studies of environmental carcinogens

BMC Cancer ◽

10.1186/s12885-021-07908-3 ◽

2021 ◽

Vol 21 (1) ◽

Author(s):

K. A. Mundt ◽

L. D. Dell ◽

P. Boffetta ◽

E. M. Beckett ◽

H. N. Lynch ◽

...

Keyword(s):

Tobacco Smoking ◽

Myeloid Leukemia ◽

Myeloproliferative Neoplasms ◽

Epidemiological Studies ◽

Sufficient Evidence ◽

International Agency ◽

Environmental Carcinogens ◽

Epidemiological Evidence ◽

Myeloid Malignancies ◽

Chemical Agents

Abstract Introduction Although myelodysplastic syndrome (MDS), acute myeloid leukemia (AML), myeloproliferative neoplasms (MPN) – including chronic myeloid leukemia (CML) – and myelodysplastic/myeloproliferative neoplasms (MDS/MPN) are largely clinically distinct myeloid malignancies, epidemiological studies rarely examine them separately and often combine them with lymphoid malignancies, limiting possible etiological interpretations for specific myeloid malignancies. Methods We systematically evaluated the epidemiological literature on the four chemical agents (1,3-butadiene, formaldehyde, benzene, and tobacco smoking, excluding pharmaceutical, microbial and radioactive agents, and pesticides) classified by the International Agency for Research on Cancer as having sufficient epidemiological evidence to conclude that each causes “myeloid malignancies.” Literature searches of IARC Monographs and PubMed identified 85 studies that we critically assessed, and for appropriate subsets, summarized results using meta-analysis. Results Only two epidemiological studies on 1,3-butadiene were identified, but reported findings were inadequate to evaluate specific myeloid malignancies. Studies on formaldehyde reported results for AML and CML – and not for MDS or MPN – but reported no increased risks. For benzene, several specific myeloid malignancies were evaluated, with consistent associations reported with AML and MDS and mixed results for CML. Studies of tobacco smoking examined all major myeloid malignancies, demonstrating consistent relationships with AML, MDS and MPN, but not with CML. Conclusions Surprisingly few epidemiological studies present results for specific myeloid malignancies, and those identified were inconsistent across studies of the same exposure, as well as across chemical agents. This exercise illustrates that even for agents classified as having sufficient evidence of causing “myeloid malignancies,” the epidemiological evidence for specific myeloid malignancies is generally limited and inconsistent. Future epidemiological studies should report findings for the specific myeloid malignancies, as combining them post hoc – where appropriate – always remains possible, whereas disaggregation may not. Furthermore, combining results across possibly discrete diseases reduces the chances of identifying important malignancy-specific causal associations.

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Persistence of minimal residual disease assessed by multiparameter flow cytometry is highly prognostic in younger patients with acute myeloid leukemia

Cancer ◽

10.1002/cncr.30361 ◽

2016 ◽

Vol 123 (3) ◽

pp. 426-435 ◽

Cited By ~ 28

Author(s):

Farhad Ravandi ◽

Jeffrey Jorgensen ◽

Gautam Borthakur ◽

Elias Jabbour ◽

Tapan Kadia ◽

...

Keyword(s):

Flow Cytometry ◽

Acute Myeloid Leukemia ◽

Minimal Residual Disease ◽

Myeloid Leukemia ◽

Residual Disease ◽

Multiparameter Flow Cytometry ◽

Younger Patients ◽

Minimal Residual ◽

Acute Myeloid

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Assessment of Immunotoxicity by Multiparameter Flow Cytometry

Toxicological Sciences ◽

10.1093/toxsci/38.1.38 ◽

1997 ◽

Vol 38 (1) ◽

pp. 38-54

Author(s):

Scott W. Burchiel ◽

Nancy L. Kerkvliet ◽

G. Frank Gerberick ◽

David A. Lawrence ◽

Gregory S. Ladics

Keyword(s):

Flow Cytometry ◽

Multiparameter Flow Cytometry

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Kinetics of virus adsorption to single cells using fluorescent membrane probes and multiparameter flow cytometry

Cell Biophysics ◽

10.1007/bf02788556 ◽

1982 ◽

Vol 4 (1) ◽

pp. 63-75 ◽

Cited By ~ 11

Author(s):

J. F. Leary ◽

M. F. D. Notter

Keyword(s):

Flow Cytometry ◽

Single Cells ◽

Multiparameter Flow Cytometry ◽

Virus Adsorption ◽

Kinetics Of ◽

Membrane Probes

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T2050 Immunophenotypic Characterization of the Cellular Infiltrate During Murine Experimental Colitis Using Multiparameter Flow Cytometry

Gastroenterology ◽

10.1016/s0016-5085(10)62863-6 ◽

2010 ◽

Vol 138 (5) ◽

pp. S-621

Author(s):

Kimberly A. Zins ◽

Tamas Ordog ◽

Michael R. Bardsley ◽

Gianrico Farrugia ◽

Joseph H. Szurszewski ◽

...

Keyword(s):

Flow Cytometry ◽

Experimental Colitis ◽

Multiparameter Flow Cytometry ◽

Cellular Infiltrate

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Immunophenotypic Study of Basophils by Multiparameter Flow Cytometry

Archives of Pathology & Laboratory Medicine ◽

10.5858/2008-132-813-isobbm ◽

2008 ◽

Vol 132 (5) ◽

pp. 813-819

Author(s):

Xiaohong Han ◽

Jeffrey L. Jorgensen ◽

Archana Brahmandam ◽

Ellen Schlette ◽

Yang O. Huh ◽

...

Keyword(s):

Bone Marrow ◽

Flow Cytometry ◽

Chronic Myelogenous Leukemia ◽

Peripheral Blood ◽

Myelogenous Leukemia ◽

Multiparameter Flow Cytometry ◽

Color Flow ◽

Aberrant Expression ◽

Flow Cytometric ◽

Hla Dr

Abstract Context.—The immunophenotypic profile of basophils is not yet fully established, and the immunophenotypic changes in chronic myelogenous leukemia are not fully characterized. Objective.—To establish a comprehensive immunophenotypic spectrum of normal basophils and to assess the range of immunophenotypic aberrations of basophils in chronic myelogenous leukemia. Design.—Using 4-color flow cytometry, we compared the immunophenotypic profile of basophils in peripheral blood or bone marrow samples from 20 patients with no evidence of neoplasia to basophils from 15 patients with chronic myelogenous leukemia. Results.—Basophils in control cases were all positive for CD9, CD13, CD22, CD25 (dim), CD33, CD36, CD38 (bright), CD45 (dimmer than lymphocytes and brighter than myeloblasts), and CD123 (bright), and were negative for CD19, CD34, CD64, CD117, and HLA-DR. Basophils in all chronic myelogenous leukemia patients possessed 1 to 5 immunophenotypic aberrancies. The most common aberrancies were underexpression of CD38, followed by aberrant expression of CD64 and underexpression of CD123. CD34 and CD117 were present in cases with basophilic precursors. Myeloblasts showed a distinct immunophenotypic profile, as they typically expressed CD34 and CD117, showed dimmer expression (compared with basophils) of CD38, CD45, and CD123, and lacked expression of CD22. Conclusions.—Flow cytometric immunophenotyping can identify immunophenotypic aberrations of basophils in chronic myelogenous leukemia, and discriminate basophils from myeloblasts.

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