Skip to main content Accessibility help

We use cookies to distinguish you from other users and to provide you with a better experience on our websites. Close this message to accept cookies or find out how to manage your cookie settings.

Close cookie message
×
Hostname: page-component-745bb68f8f-kw2vx Total loading time: 0 Render date: 2025-01-31T23:41:10.872Z Has data issue: false hasContentIssue false

Chapter 4 - Reactive Conditions and Other Diseases Where Flow Cytometric Findings May Mimic Haematological Malignancies

Published online by Cambridge University Press:  30 January 2025

By
Marie-Christine Béné ,
Wolfgang Kern  and
Anna Porwit
Edited by
Anna Porwit  and
Marie Christine Béné
Anna Porwit
Affiliation:
Lunds Universitet, Sweden
Marie Christine Béné
Affiliation:
Université de Nantes, France
Get access

Summary

As indicated by the title of this chapter, some conditions may display features evocative of hematological malignancies and have to be recognized as not being tumoral. Here, these situations have been grouped as increased leucocyte types (leucocytosis) or as decreased cell counts (cytopenias), segregated in disease types. A third part considers abnormal immunophenotypes of lymphocytes and myeloid cells. Finally, the recurrent question of haemodilution of bone marrow aspirates, which decreases the otherwise helpful ability of flow cytometry to count large numbers of cells and thus perform accurate differentials, is discussed.

Keywords

flow cytometrynon-malignant conditionslymphocytosisleucocytosismyeloid cellsmonocytosiscytopeniainfectionabnormal immunophenotypeshaemodilution
Type
Chapter
Information
Multiparameter Flow Cytometry in the Diagnosis of Hematologic Malignancies , pp. 59 - 72
Publisher: Cambridge University Press
Print publication year: 2025

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Craig, FE, Foon, KA. Flow cytometric immunophenotyping for hematologic neoplasms. Blood 2008; 111: 3941–3967.CrossRefGoogle ScholarPubMed
Davis, BH, Holden, JT, Bene, MC, et al. 2006 Bethesda International Consensus recommendations on the flow cytometric immunophenotypic analysis of hematolymphoid neoplasia: medical indications. Cytometry B Clin Cytom 2007; 72: S5–S13.CrossRefGoogle ScholarPubMed
Johansson, U, Bloxham, D, Couzens, S, et al.; British Committee for Standards in Haematology. Guidelines on the use of multicolour flow cytometry in the diagnosis of haematological neoplasms. Br J Haematol 2014; 165: 455–488.CrossRefGoogle ScholarPubMed
Arber, DA, Orazi, A, Hasserjian, R, et al. The 2016 revision to the World Health Organization classification of myeloid neoplasms and acute leukemia. Blood 2016; 127: 2391–2405.CrossRefGoogle Scholar
Kern, W, Haferlach, C, Schnittger, S, Haferlach, T. Clinical utility of multiparameter flow cytometry in the diagnosis of 1013 patients with suspected myelodysplastic syndrome: correlation to cytomorphology, cytogenetics, and clinical data. Cancer 2010; 116: 4549–4563.CrossRefGoogle ScholarPubMed
Morizot, R, de Korwin, JD, Feugier, P, Broséus, J, Troussard, X, Lesesve, JF. Patients with persistent polyclonal b-cell lymphocytosis share the symptomatic criteria of systemic exertion intolerance disease. J Clin Med 2021; 10: 3374CrossRefGoogle ScholarPubMed
Himmelmann, A, Gautschi, O, Nawrath, M, Bolliger, U, Fehr, J, Stahel, RA. Persistent polyclonal B-cell lymphocytosis is an expansion of functional IgD(+)CD27(+) memory B cells. Br J Haematol 2001; 114: 400–405.CrossRefGoogle Scholar
Cornet, E, Mossafa, H, Courel, K, Lesesve, JF, Troussard, X. Persistent polyclonal binucleated B-cell lymphocytosis and MECOM gene amplification. BMC Res Notes 2016; 9: 138.CrossRefGoogle ScholarPubMed
Tesson, B, Huet, S, Grange, B, et al. Absence of driver mutations in persistent polyclonal B-cell lymphocytosis with binucleated lymphocytes. Blood 2017; 130: 1267–1269.CrossRefGoogle ScholarPubMed
Mossafa, H, Malaure, H, Maynadie, M, et al. Persistent polyclonal B lymphocytosis with binucleated lymphocytes: a study of 25 cases. Groupe Français d’Hématologie Cellulaire. Br J Haematol 1999; 104: 486–493.CrossRefGoogle ScholarPubMed
Zhao, Y, Siddiqi, I, Wildes, TJ, et al. Chronic lymphocytic leukemia with two B-cell populations of discordant light chain restrictions in individual patients: parallel development of biclonal B-cell neoplasms or clonal evolution with isotype switch? Am J Clin Pathol 2023; 159: 337–351.CrossRefGoogle ScholarPubMed
Mahdi, T, Rajab, A, Padmore, R, Porwit, A. Characteristics of lymphoproliferative disorders with more than one aberrant cell population as detected by 10-color flow cytometry. Cytometry B Clin Cytom 2018; 94: 230–238.CrossRefGoogle ScholarPubMed
Lima, M, Teixeira Mdos, A, Queirós, ML, et al. Immunophenotype and TCR-Vbeta repertoire of peripheral blood T-cells in acute infectious mononucleosis. Blood Cells Mol Dis 2003; 30: 1–12.CrossRefGoogle ScholarPubMed
Strioga, M, Pasukoniene, V, Characiejus, D. CD8+ CD28- and CD8+ CD57+ T cells and their role in health and disease. Immunology 2011; 134: 17–32.CrossRefGoogle ScholarPubMed
Moon, Y, Jang, WR, Yi, HG, et al. Klebsiella pneumoniae associated extreme plasmacytosis. Infect Chemother 2013; 45: 435–440.CrossRefGoogle ScholarPubMed
Gawoski, JM, Ooi, WW. Dengue fever mimicking plasma cell leukemia. Arch Pathol Lab Med 2003; 127: 1026–1027.CrossRefGoogle ScholarPubMed
Kumar, S, Kimlinger, T, Morice, W. Immunophenotyping in multiple myeloma and related plasma cell disorders. Best Pract Res Clin Haematol 2010; 23: 433–451.CrossRefGoogle ScholarPubMed
Davis, BH, Olsen, SH, Ahmad, E, Bigelow, NC. Neutrophil CD64 is an improved indicator of infection or sepsis in emergency department patients. Arch Pathol Lab Med 2006; 130: 654–661.CrossRefGoogle ScholarPubMed
Ouyang, J, Zheng, W, Shen, Q, et al. Flow cytometry immunophenotypic analysis of Philadelphia-negative myeloproliferative neoplasms: correlation with histopathologic features. Cytometry B Clin Cytom 2015; 88: 236–243.CrossRefGoogle ScholarPubMed
Mangaonkar, AA, Tande, AJ, Bekele, DI. Differential diagnosis and workup of monocytosis: a systematic approach to a common hematologic finding. Curr Hematol Malig Rep. 2021; 16: 267–270.CrossRefGoogle ScholarPubMed
Schulte-Schrepping, J, Reusch, N, Paclik, D, et al. Severe COVID-19 is marked by a dysregulated myeloid cell compartment. Cell. 2020; 182: 1419–1440.CrossRefGoogle ScholarPubMed
Selimoglu-Buet, D, Wagner-Ballon, O, Saada, V, et al. Characteristic repartition of monocyte subsets as a diagnostic signature of chronic myelomonocytic leukemia. Blood. 2015; 125: 3618–3626.CrossRefGoogle ScholarPubMed
Arnoulet, C, Béné, MC, Durrieu, F, et al. Four- and five-color flow cytometry analysis of leukocyte differentiation pathways in normal bone marrow: a reference document based on a systematic approach by the GTLLF and GEIL. Cytometry B Clin Cytom 2010; 78: 4–10.CrossRefGoogle ScholarPubMed
Orazi, A. Histopathology in the diagnosis and classification of acute myeloid leukemia, myelodysplastic syndromes, and myelodysplastic/myeloproliferative diseases. Pathobiology 2007; 74: 97–114.CrossRefGoogle ScholarPubMed
Porwit, A, Fend, F, Kremer, M, Orazi, A, Safali, M, van der Walt, J. Issues in diagnosis of small B cell lymphoid neoplasms involving the bone marrow and peripheral blood: report on the Bone Marrow Workshop of the XVIIth meeting of the European Association for Haematopathology and the Society for Hematopathology. Histopathology 2016; 69: 349–373.CrossRefGoogle Scholar
Maciejewski, JP, Hibbs, JR, Anderson, S, Katevas, P, Young, NS. Bone marrow and peripheral blood lymphocyte phenotype in patients with bone marrow failure. Exp Hematol 1994; 22: 1102–1110.Google ScholarPubMed
Risitano, AM, Kook, H, Zeng, W, Chen, G, Young, NS, Maciejewski, JP. Oligoclonal and polyclonal CD4 and CD8 lymphocytes in aplastic anemia and paroxysmal nocturnal hemoglobinuria measured by V beta CDR3 spectratyping and flow cytometry. Blood 2002; 100: 178–183.CrossRefGoogle ScholarPubMed
Lu, T, Liu, Y, Li, P, et al. Decreased circulating Th22 and Th17 cells in patients with aplastic anemia. Clin Chim Acta 2015; 450: 90–96.CrossRefGoogle ScholarPubMed
de Latour, RP, Visconte, V, Takaku, T, et al. Th17 immune responses contribute to the pathophysiology of aplastic anemia. Blood 2010; 116: 4175–4184.CrossRefGoogle Scholar
Manz, CY, Nissen, C, Wodnar-Filipowicz, A. Deficiency of CD34+ c-kit+ and CD34+38- hematopoietic precursors in aplastic anemia after immunosuppressive treatment. Am J Hematol 1996; 52: 264–274.3.0.CO;2-Q>CrossRefGoogle ScholarPubMed
Maciejewski, JP, Anderson, S, Katevas, P, Young, NS. Phenotypic and functional analysis of bone marrow progenitor cell compartment in bone marrow failure. Br J Haematol 1994; 87: 227–234.CrossRefGoogle ScholarPubMed
Huang, M, Li, J, Zhao, G, et al. Immunophenotype of myeloid granulocytes: a pilot study for distinguishing myelodysplastic syndrome and aplastic anemia by flow cytometry. Int J Lab Hematol 2010; 32: 275–281.CrossRefGoogle ScholarPubMed
Sevilla, DW, Emmons, FN, Colovai, AI, et al. Hematogones are markedly reduced in pediatric acquired aplastic anemia: multiparametric flow cytometric analysis. Leuk Lymphoma 2009; 50: 1951–1957.CrossRefGoogle ScholarPubMed
Young, NS, Maciejewski, JP, Sloand, E, et al. The relationship of aplastic anemia and PNH. Int J Hematol 2002; 76: 168–172.CrossRefGoogle ScholarPubMed
Borowitz, MJ, Craig, FE, Digiuseppe, JA, et al.; Clinical Cytometry Society. Guidelines for the diagnosis and monitoring of paroxysmal nocturnal hemoglobinuria and related disorders by flow cytometry. Cytometry B Clin Cytom 2010; 78: 211–230.CrossRefGoogle ScholarPubMed
Sutherland, DR, Keeney, M, Illingworth, A. Practical guidelines for the high-sensitivity detection and monitoring of paroxysmal nocturnal hemoglobinuria clones by flow cytometry. Cytometry B Clin Cytom 2012; 82: 195–208.CrossRefGoogle ScholarPubMed
Illingworth, AJ, Marinov, I, Sutherland, DR. Sensitive and accurate identification of PNH clones based on ICCS/ESCCA PNH Consensus Guidelines: a summary. Int J Lab Hematol 2019; 41: 73–81.CrossRefGoogle ScholarPubMed
Raza, A, Ravandi, F, Rastogi, A, et al. A prospective multicenter study of paroxysmal nocturnal hemoglobinuria cells in patients with bone marrow failure. Cytometry B Clin Cytom 2014; 86: 175–182.CrossRefGoogle ScholarPubMed
Muroi, K, Fujiwara, S, Tatara, R, et al. CD56 expression in normal immature granulocytes after allogeneic hematopoietic stem cell transplantation. J Clin Exp Hematop 2013; 53: 247–250.CrossRefGoogle ScholarPubMed
Xu, Y, McKenna, RW, Karandikar, NJ, Pildain, AJ, Kroft, SH. Flow cytometric analysis of monocytes as a tool for distinguishing chronic myelomonocytic leukemia from reactive monocytosis. Am J Clin Pathol 2005; 124: 799–806.CrossRefGoogle ScholarPubMed
Kern, W, Danhauser-Riedl, S, Ratei, R, et al. Detection of minimal residual disease in unselected patients with acute myeloid leukemia using multiparameter flow cytometry for definition of leukemia-associated immunophenotypes and determination of their frequencies in normal bone marrow. Haematologica 2003; 88: 646–653.Google ScholarPubMed
Curtis, BR. Drug-induced immune neutropenia/agranulocytosis. Immunohematology 2014; 30: 95–101.CrossRefGoogle ScholarPubMed
Bourré-Tessier, J, Haraoui, B. Methotrexate drug interactions in the treatment of rheumatoid arthritis: a systematic review. J Rheumatol 2010; 37: 1416–1421.CrossRefGoogle ScholarPubMed
Curtis, BR. Drug-induced immune thrombocytopenia: incidence, clinical features, laboratory testing, and pathogenic mechanisms. Immunohematology 2014; 30: 55–65.CrossRefGoogle ScholarPubMed
Tomer, A. Flow cytometry for the diagnosis of autoimmune thrombocytopenia. Curr Hematol Rep 2006; 5: 64–69.Google ScholarPubMed
Zhao, Y, Zhu, M, Jiang, M, et al. An improved flow cytometric immunobead array to detect autoantibodies in plasma from patients with immune thrombocytopenic purpura. Clin Chim Acta 2015; 438: 396–400.CrossRefGoogle ScholarPubMed
Porwit, A, Violidaki, D, Axler, O, Lacombe, F, Ehinger, M, Béné, MC. Unsupervised cluster analysis and subset characterization of abnormal erythropoiesis using the bioinformatic Flow-Self Organizing Maps algorithm. Cytometry B Clin Cytom 2022; 102: 134–142.CrossRefGoogle ScholarPubMed
King, MJ, Behrens, J, Rogers, C, Flynn, C, Greenwood, D, Chambers, K. Rapid flow cytometric test for the diagnosis of membrane cytoskeleton-associated haemolytic anaemia. Br J Haematol 2000; 111: 924–933.Google ScholarPubMed
Singleton, BK, Ahmed, M, Green, CA, et al. CD44 as a potential screening marker for preliminary differentiation between congenital dyserythropoietic anemia type II and hereditary spherocytosis. Cytometry B Clin Cytom 2018; 94: 312–326.CrossRefGoogle ScholarPubMed
Thedsawad, A, Taka, O, Wanachiwanawin, W. Significances of red cell bound immunoglobulin G as detected by flow cytometry in patients with Coombs-negative immune hemolysis. Transfus Med 2016; 26: 130–137.CrossRefGoogle ScholarPubMed
Steensma, DP. Dysplasia has a differential diagnosis: distinguishing genuine myelodysplastic syndromes (MDS) from mimics, imitators, copycats and impostors. Curr Hematol Malig Rep 2012; 7: 310–320.CrossRefGoogle Scholar
Malcovati, L, Cazzola, M. The shadowlands of MDS: idiopathic cytopenias of undetermined significance (ICUS) and clonal hematopoiesis of indeterminate potential (CHIP). Hematology Am Soc Hematol Educ Program 2015; 2015: 299–307.CrossRefGoogle ScholarPubMed
Steensma, DP, Bejar, R, Jaiswal, S, et al. Clonal hematopoiesis of indeterminate potential and its distinction from myelodysplastic syndromes. Blood 2015; 126: 9–16.CrossRefGoogle ScholarPubMed
Therkelsen, J, Traeden, DW, Schjødt, I, et al. ProGraME: A novel flow cytometry algorithm for the diagnosis of low-risk myelodysplastic syndromes in patients with cytopenia. Eur J Haematol 2023; 111: 851–862.CrossRefGoogle ScholarPubMed
Cossarizza, A, De Biasi, S, Gibellini, L, Bianchini, E, Bartolomeo, R, Nasi, M, Mussini, C, Pinti, M. Cytometry, immunology, and HIV infection: three decades of strong interactions. Cytometry A 2013; 83: 680–691.CrossRefGoogle ScholarPubMed
Ron, R, Moreno, E, Martínez-Sanz, J, et al. CD4/CD8 ratio during human immunodeficiency virus treatment: time for routine monitoring? Clin Infect Dis 2023; 76: 1688–1696.CrossRefGoogle ScholarPubMed
Hagen, M, Pangrazzi, L, Rocamora-Reverte, L, Weinberger, B. Legend or truth: mature CD4+CD8+ double-positive T cells in the periphery in health and disease. Biomedicines 2023; 11: 2702.CrossRefGoogle ScholarPubMed
Chen, Z, John, Wherry E. T cell responses in patients with COVID-19. Nat Rev Immunol 2020; 20: 529–536.CrossRefGoogle ScholarPubMed
Du, J, Wei, L, Li, G, et al. Persistent high percentage of HLA-DR+CD38high CD8+ T cells associated with immune disorder and disease severity of COVID-19. Front Immunol 2021; 12: 735125.CrossRefGoogle ScholarPubMed
Ouyang, J, Zheng, W, Shen, Q, et al. Flow cytometry immunophenotypic analysis of Philadelphia-negative myeloproliferative neoplasms: correlation with histopathologic features. Cytometry B Clin Cytom 2015; 88: 236–243.CrossRefGoogle ScholarPubMed
Loken, MR, Chu, SC, Fritschle, W, Kalnoski, M, Wells, DA. Normalization of bone marrow aspirates for hemodilution in flow cytometric analyses. Cytometry B Clin Cytom 2009; 76: 27–36.CrossRefGoogle ScholarPubMed
Aldawood, AM, Kinkade, Z, Rosado, FG, Esan, OA, Gibson, LF, Vos, JA. A novel method to assess bone marrow purity is useful in determining blast percentage by flow cytometry in acute myeloid leukemia and myelodysplasia. Ann Hematol Oncol 2015; 2: 1038.Google ScholarPubMed
Delgado, JA, Guillén-Grima, F, Moreno, C, et al. A simple flow-cytometry method to evaluate peripheral blood contamination of bone marrow aspirates. J Immunol Methods 2017; 442: 54–58.CrossRefGoogle ScholarPubMed
Holdrinet, RS, von Egmond, J, Wessels, JM, et al. A method for quantification of peripheral blood admixture in bone marrow aspirates. Exp Hematol 1980; 8: 103–107.Google ScholarPubMed
Flores-Montero, J, Sanoja-Flores, L, Paiva, B, et al. Next Generation Flow for highly sensitive and standardized detection of minimal residual disease in multiple myeloma. Leukemia 2017; 31: 2094–2103.CrossRefGoogle ScholarPubMed
Tettero, JM, Freeman, S, Buecklein, V, et al. Technical aspects of flow cytometry-based measurable residual disease quantification in acute myeloid leukemia: experience of the European LeukemiaNet MRD Working Party. Hemasphere 2021; 22: e676.Google Scholar

Save book to Kindle

To save this book to your Kindle, first ensure no-reply@cambridge.org is added to your Approved Personal Document E-mail List under your Personal Document Settings on the Manage Your Content and Devices page of your Amazon account. Then enter the ‘name’ part of your Kindle email address below. Find out more about saving to your Kindle.

Note you can select to save to either the @free.kindle.com or @kindle.com variations. ‘@free.kindle.com’ emails are free but can only be saved to your device when it is connected to wi-fi. ‘@kindle.com’ emails can be delivered even when you are not connected to wi-fi, but note that service fees apply.

Find out more about the Kindle Personal Document Service.

Available formats
×

Save book to Dropbox

To save content items to your account, please confirm that you agree to abide by our usage policies. If this is the first time you use this feature, you will be asked to authorise Cambridge Core to connect with your account. Find out more about saving content to Dropbox.

Available formats
×

Save book to Google Drive

To save content items to your account, please confirm that you agree to abide by our usage policies. If this is the first time you use this feature, you will be asked to authorise Cambridge Core to connect with your account. Find out more about saving content to Google Drive.

Available formats
×