CC BY-NC-ND 4.0 · J Lab Physicians 2023; 15(03): 392-398
DOI: 10.1055/s-0043-1761204
Original Article

Immunophenotypic Profile of Multiple Myeloma: A Tertiary Care Centre Experience

1   Department of Hematology, All India Institute of Medical Sciences, New Delhi, India
Tribikram Panda
1   Department of Hematology, All India Institute of Medical Sciences, New Delhi, India
Jasmita Dass
1   Department of Hematology, All India Institute of Medical Sciences, New Delhi, India
Tulika Seth
1   Department of Hematology, All India Institute of Medical Sciences, New Delhi, India
Manoranjan Mahapatra
1   Department of Hematology, All India Institute of Medical Sciences, New Delhi, India
Seema Tyagi
1   Department of Hematology, All India Institute of Medical Sciences, New Delhi, India
› Author Affiliations
Funding None.


Background Immunophenotyping and enumeration of plasma cells (PCs) by flow cytometry are deemed to be prognostically significant. However, PCs enumeration by flow cytometry is challenging owing to discrepancy with morphology and PCs loss during sample processing. Enumeration and differentiation of abnormal plasma cells (APCs) and normal plasma cells (NPCs) is difficult because abnormal antigen expression can be seen in subsets of NPCs. This is particularly true when a limited panel of antibodies are relied upon.

Aims and purpose To study the immunophenotypic profile of newly diagnosed multiple myeloma (MM) cases by flow cytometry and evaluate the sensitivities and specificities of individual antigens and combinations.

Methods We studied immunophenotype of PCs in newly diagnosed MM cases (n = 48) and control cases (n = 10) by a 6-color, 3-tube flow cytometry panel. The sensitivities and specificities of antigens in MM were evaluated and compared with control cases.

Results Majority of MM cases (n = 43) had < 3% NPCs. CD19 was the most sensitive (100%) and CD81 was the most specific marker (100%) for differentiating APCs from NPCs. CD38 MFI came out as a useful marker for APCs identification. In combination, CD19 and CD81 had a higher sensitivity and specificity to detect APCs.

Conclusion NPCs may show aberrant antigen expression. A combination of multiple markers including CD81 and CD38 MFI should be used for accurate APC detection.

Ethical Standards

All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki Declaration and its later amendments or comparable ethical standards.

Ethical Approval

The study is approved by institute ethics committee with reference number IECPG-293/29.05.2019.

Informed Consent

Informed consent was obtained from all individual participants included in the study.

Publication Consent

Consent for publication was obtained for every individual person's data included in the study.

Data Availability

The datasets generated and analyzed during this study are available from the corresponding author on reasonable request.

Authors Contributions

A.R. processed the samples, analyzed the flow cytometry data, and wrote the manuscript. S.T. conceptualized and designed the study, analyzed the data, and wrote the manuscript. J.D. reviewed the morphology and analyzed the data. T.S. and M.M. reviewed the data and provided the clinical information.

Publication History

Article published online:
30 January 2023

© 2023. The Indian Association of Laboratory Physicians. This is an open access article published by Thieme under the terms of the Creative Commons Attribution-NonDerivative-NonCommercial License, permitting copying and reproduction so long as the original work is given appropriate credit. Contents may not be used for commercial purposes, or adapted, remixed, transformed or built upon. (

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  • References

  • 1 McKenna R, Kyle R, Kuehl W. Plasma cell neoplasms. In: WHO classification of tumours of haematopoietic and lymphoid tissues. revised 4th ed. 2017 Lyon; IACR:241–58
  • 2 Rajkumar SV, Dimopoulos MA, Palumbo A. et al. International Myeloma Working Group updated criteria for the diagnosis of multiple myeloma. Lancet Oncol 2014; 15 (12) e538-e548
  • 3 Jelinek T, Bezdekova R, Zatopkova M. et al. Current applications of multiparameter flow cytometry in plasma cell disorders. Blood Cancer J 2017; 7 (10) e617-e617
  • 4 San Miguel JF, Almeida J, Mateo G. et al. Immunophenotypic evaluation of the plasma cell compartment in multiple myeloma: a tool for comparing the efficacy of different treatment strategies and predicting outcome. Blood 2002; 99 (05) 1853-1856
  • 5 Sezer O, Heider U, Zavrski I, Possinger K. Differentiation of monoclonal gammopathy of undetermined significance and multiple myeloma using flow cytometric characteristics of plasma cells. Haematologica 2001; 86 (08) 837-843
  • 6 Pérez-Persona E, Vidriales M-B, Mateo G. et al. New criteria to identify risk of progression in monoclonal gammopathy of uncertain significance and smoldering multiple myeloma based on multiparameter flow cytometry analysis of bone marrow plasma cells. Blood 2007; 110 (07) 2586-2592
  • 7 Paiva B, Vidriales M-B, Mateo G. et al; GEM (Grupo Español de MM)/PETHEMA (Programa para el Estudio de la Terapéutica en Hemopatías Malignas) Cooperative Study Groups. The persistence of immunophenotypically normal residual bone marrow plasma cells at diagnosis identifies a good prognostic subgroup of symptomatic multiple myeloma patients. Blood 2009; 114 (20) 4369-4372
  • 8 Kumar S, Kimlinger T, Morice W. Immunophenotyping in multiple myeloma and related plasma cell disorders. Best Pract Res Clin Haematol 2010; 23 (03) 433-451
  • 9 Tembhare PR, Yuan CM, Venzon D. et al. Flow cytometric differentiation of abnormal and normal plasma cells in the bone marrow in patients with multiple myeloma and its precursor diseases. Leuk Res 2014; 38 (03) 371-376
  • 10 Perez-Andres M, Santiago M, Almeida J, Mateo G, Porwit-Macdonald A, Bjorklund E, Valet G, Kraan J, Gratama J, D'Hautcourt JL, Merle-Beral H. Immunophenotypic approach to the identification and Journal of Biological Regulators and Homeostatic Agents. 2004
  • 11 Rawstron AC, Orfao A, Beksac M. et al; European Myeloma Network. Report of the European Myeloma Network on multiparametric flow cytometry in multiple myeloma and related disorders. Haematologica 2008; 93 (03) 431-438
  • 12 Nadav L, Katz BZ, Baron S. et al. Diverse niches within multiple myeloma bone marrow aspirates affect plasma cell enumeration. Br J Haematol 2006; 133 (05) 530-532
  • 13 Manasanch EE, Salem DA, Yuan CM. et al. Flow cytometric sensitivity and characteristics of plasma cells in patients with multiple myeloma or its precursor disease: influence of biopsy site and anticoagulation method. Leuk Lymphoma 2015; 56 (05) 1416-1424
  • 14 Lu J, Li C, Huang Y, Zhang J. Comparison of cell morphology and flow cytometry in the diagnosis of multiple myeloma. J Cancer Ther 2020; 11 (11) 731-737
  • 15 Bacher U, Haferlach T, Kern W, Alpermann T, Schnittger S, Haferlach C. Correlation of cytomorphology, immunophenotyping, and interphase fluorescence in situ hybridization in 381 patients with monoclonal gammopathy of undetermined significance and 301 patients with plasma cell myeloma. Cancer Genet Cytogenet 2010; 203 (02) 169-175
  • 16 Tran DN, Smith SABC, Brown DA. et al. Polychromatic flow cytometry is more sensitive than microscopy in detecting small monoclonal plasma cell populations. Cytometry B Clin Cytom 2017; 92 (02) 136-144
  • 17 Kalina T, Flores-Montero J, van der Velden VHJ. et al; EuroFlow Consortium (EU-FP6, LSHB-CT-2006-018708). EuroFlow standardization of flow cytometer instrument settings and immunophenotyping protocols. Leukemia 2012; 26 (09) 1986-2010
  • 18 Chatterjee G, Gujral S, Subramanian PG, Tembhare PR. Clinical relevance of multicolour flow cytometry in plasma cell disorders. Indian J Hematol Blood Transfus 2017; 33 (03) 303-315
  • 19 Ocqueteau M, Orfao A, Almeida J. et al. Immunophenotypic characterization of plasma cells from monoclonal gammopathy of undetermined significance patients. Implications for the differential diagnosis between MGUS and multiple myeloma. Am J Pathol 1998; 152 (06) 1655-1665
  • 20 Paiva B, Almeida J, Pérez-Andrés M. et al. Utility of flow cytometry immunophenotyping in multiple myeloma and other clonal plasma cell-related disorders. Cytometry B Clin Cytom 2010; 78 (04) 239-252
  • 21 Peceliunas V, Janiulioniene A, Matuzeviciene R, Griskevicius L. Six color flow cytometry detects plasma cells expressing aberrant immunophenotype in bone marrow of healthy donors. Cytometry B Clin Cytom 2011; 80 (05) 318-323
  • 22 Pojero F, Flores-Montero J, Sanoja L. et al; EuroFlow group. Utility of CD54, CD229, and CD319 for the identification of plasma cells in patients with clonal plasma cell diseases. Cytometry B Clin Cytom 2016; 90 (01) 91-100