Flow Cytometric Evaluation of Traditional and Novel Surface Markers for the Diagnosis of Plasma Cell Dyscrasias

Abstract

Increasing interest has been expressed for flow cytometric immunophenotyping for diagnosis and monitoring in plasma cell dyscrasias over the last decades. The aim of this investigation was to compare the expression strength of various cell surface markers used traditionally or currently under investigation on normal and abnormal PC populations. We enrolled 295 consecutive patients undergoing bone marrow aspiration in the workup of monoclonal gammopathies, selecting 54 normal and 241 abnormal PC populations via flow cytometry to characterize the expression of CD45, CD38, CD138, CD19, CD56, CD20, CD27, CD28, CD81, CD117 and CD200 on the cell surface of PCs. We observed significant differences in the expression strength of all assessed markers between normal and abnormal PC populations in all markers except for CD20. While none of them was conclusive on its own, the combination of CD81 positivity and CD117 negativity was present in 98.1% of normal PC populations tested. In contrast, particularly CD117 positivity, but also CD81 negativity was indicative of an abnormal PC phenotype. Our results highlight the descriptive value of CD81 and CD117 for the allocation of bone marrow PCs to a normal or abnormal phenotype.

This is a preview of subscription content, log in to check access.

Fig. 1
Fig. 2

Abbreviations

mAbs:

Monoclonal antibodies

MGUS:

Monoclonal gammopathy of undetermined significance

PC:

Plasma cell

NK cell:

Natural killer cell

References

  1. 1.

    Kyle RA, Rajkumar SV (2004) Multiple myeloma. N Engl J Med 18:1860–1873

    Article  Google Scholar 

  2. 2.

    Rajkumar SV (2013) Multiple myeloma: 2013 update on diagnosis, risk-stratification, and management. Am J Hematol 3:226–235

    Google Scholar 

  3. 3.

    Chesi M, Bergsagel PL (2013) Molecular pathogenesis of multiple myeloma: basic and clinical updates. Int J Hematol 3:313–323

    Article  CAS  Google Scholar 

  4. 4.

    Raja KR, Kovarova L, Hajek R (2010) Review of phenotypic markers used in flow cytometric analysis of MGUS and MM, and applicability of flow cytometry in other plasma cell disorders. Br J Haematol 3:334–351

    Article  CAS  Google Scholar 

  5. 5.

    Palumbo A, Anderson K (2011) Multiple myeloma. N Engl J Med 11:1046–1060

    Article  Google Scholar 

  6. 6.

    Kyle RA, Rajkumar SV (2008) Multiple myeloma. Blood 6:2962–2972

    Article  Google Scholar 

  7. 7.

    Raab MS, Podar K, Breitkreutz I, Richardson PG, Anderson KC (2009) Multiple myeloma. Lancet 9686:324–339

    Article  Google Scholar 

  8. 8.

    Willrich MA, Katzmann JA (2016) Laboratory testing requirements for diagnosis and follow-up of multiple myeloma and related plasma cell dyscrasias. Clin Chem Lab Med 6:907–919

    Google Scholar 

  9. 9.

    Paiva B, Almeida J, Perez-Andres M, Mateo G, Lopez A, Rasillo A et al (2010) Utility of flow cytometry immunophenotyping in multiple myeloma and other clonal plasma cell-related disorders. Cytom B Clin Cytom 4:239–252

    Google Scholar 

  10. 10.

    Tran DN, Smith SA, Brown DA, Parker AJ, Joseph JE, Armstrong N et al (2017) Polychromatic flow cytometry is more sensitive than microscopy in detecting small monoclonal plasma cell populations. Cytom B Clin Cytom 2:136–144

    Article  CAS  Google Scholar 

  11. 11.

    Rawstron AC, Orfao A, Beksac M, Bezdickova L, Brooimans RA, Bumbea H et al (2008) Report of the European Myeloma Network on multiparametric flow cytometry in multiple myeloma and related disorders. Haematologica 3:431–438

    Article  Google Scholar 

  12. 12.

    Paiva B, Vidriales MB, Rosinol L, Martinez-Lopez J, Mateos MV, Ocio EM et al (2013) A multiparameter flow cytometry immunophenotypic algorithm for the identification of newly diagnosed symptomatic myeloma with an MGUS-like signature and long-term disease control. Leukemia 10:2056–2061

    Article  Google Scholar 

  13. 13.

    Ocqueteau M, Orfao A, Almeida J, Blade J, Gonzalez M, Garcia-Sanz R et al (1998) 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 6:1655–1665

    Google Scholar 

  14. 14.

    Kobayashi S, Hyo R, Amitani Y, Tanaka M, Hashimoto C, Sakai R et al (2006) Four-color flow cytometric analysis of myeloma plasma cells. Am J Clin Pathol 6:908–915

    Article  Google Scholar 

  15. 15.

    Harada H, Kawano MM, Huang N, Harada Y, Iwato K, Tanabe O et al (1993) Phenotypic difference of normal plasma cells from mature myeloma cells. Blood 10:2658–2663

    Article  Google Scholar 

  16. 16.

    Robillard N, Avet-Loiseau H, Garand R, Moreau P, Pineau D, Rapp MJ et al (2003) CD20 is associated with a small mature plasma cell morphology and t(11;14) in multiple myeloma. Blood 3:1070–1071

    Article  CAS  Google Scholar 

  17. 17.

    Zandecki M, Facon T, Bernardi F, Izydorczyk V, Dupond L, Francois M et al (1995) CD19 and immunophenotype of bone marrow plasma cells in monoclonal gammopathy of undetermined significance. J Clin Pathol 6:548–552

    Article  Google Scholar 

  18. 18.

    Guikema JE, Hovenga S, Vellenga E, Conradie JJ, Abdulahad WH, Bekkema R et al (2003) CD27 is heterogeneously expressed in multiple myeloma: low CD27 expression in patients with high-risk disease. Br J Haematol 1:36–43

    Article  Google Scholar 

  19. 19.

    Katayama Y, Sakai A, Oue N, Asaoku H, Otsuki T, Shiomomura T et al (2003) A possible role for the loss of CD27–CD70 interaction in myelomagenesis. Br J Haematol 2:223–234

    Article  Google Scholar 

  20. 20.

    Pellat-Deceunynck C, Bataille R, Robillard N, Harousseau JL, Rapp MJ, Juge-Morineau N et al (1994) Expression of CD28 and CD40 in human myeloma cells: a comparative study with normal plasma cells. Blood 8:2597–2603

    Article  Google Scholar 

  21. 21.

    Robillard N, Jego G, Pellat-Deceunynck C, Pineau D, Puthier D, Mellerin MP et al (1998) CD28, a marker associated with tumoral expansion in multiple myeloma. Clin Cancer Res 6:1521–1526

    Google Scholar 

  22. 22.

    Ocqueteau M, Orfao A, Garcia-Sanz R, Almeida J, Gonzalez M, San Miguel JF (1996) Expression of the CD117 antigen (c-Kit) on normal and myelomatous plasma cells. Br J Haematol 3:489–493

    Article  Google Scholar 

  23. 23.

    Bataille R, Pellat-Deceunynck C, Robillard N, Avet-Loiseau H, Harousseau JL, Moreau P (2008) CD117 (c-kit) is aberrantly expressed in a subset of MGUS and multiple myeloma with unexpectedly good prognosis. Leuk Res 3:379–382

    Article  CAS  Google Scholar 

  24. 24.

    Lin P, Owens R, Tricot G, Wilson CS (2004) Flow cytometric immunophenotypic analysis of 306 cases of multiple myeloma. Am J Clin Pathol 4:482–488

    Article  Google Scholar 

  25. 25.

    Olteanu H, Harrington AM, Hari P, Kroft SH (2011) CD200 expression in plasma cell myeloma. Br J Haematol 3:408–411

    Article  Google Scholar 

  26. 26.

    Paiva B, Gutierrez NC, Chen X, Vidriales MB, Montalban MA, Rosinol L et al (2012) Clinical significance of CD81 expression by clonal plasma cells in high-risk smoldering and symptomatic multiple myeloma patients. Leukemia 8:1862–1869

    Article  CAS  Google Scholar 

  27. 27.

    Flores-Montero J, de Tute R, Paiva B, Perez JJ, Bottcher S, Wind H et al (2016) Immunophenotype of normal vs. myeloma plasma cells: toward antibody panel specifications for MRD detection in multiple myeloma. Cytom B Clin Cytom 1:61–72

    Article  CAS  Google Scholar 

  28. 28.

    Porwit A (2013) Immunophenotyping of selected hematologic disorders–focus on lymphoproliferative disorders with more than one malignant cell population. Int J Lab Hematol. 3:275–282

    Article  Google Scholar 

  29. 29.

    Cannizzo E, Carulli G, Del Vecchio L, Ottaviano V, Bellio E, Zenari E et al (2012) The role of CD19 and CD27 in the diagnosis of multiple myeloma by flow cytometry: a new statistical model. Am J Clin Pathol 3:377–386

    Article  Google Scholar 

  30. 30.

    Flanders A, Stetler-Stevenson M, Landgren O (2013) Minimal residual disease testing in multiple myeloma by flow cytometry: major heterogeneity. Blood 6:1088–1089

    Article  Google Scholar 

  31. 31.

    Bataille R, Jego G, Robillard N, Barille-Nion S, Harousseau JL, Moreau P et al (2006) The phenotype of normal, reactive and malignant plasma cells. Identification of “many and multiple myelomas” and of new targets for myeloma therapy. Haematologica 9:1234–1240

    Google Scholar 

  32. 32.

    Pellat-Deceunynck C, Bataille R (2004) Normal and malignant human plasma cells: proliferation, differentiation, and expansions in relation to CD45 expression. Blood Cells Mol Dis 2:293–301

    Article  CAS  Google Scholar 

  33. 33.

    Robillard N, Pellat-Deceunynck C, Bataille R (2005) Phenotypic characterization of the human myeloma cell growth fraction. Blood 12:4845–4848

    Article  CAS  Google Scholar 

  34. 34.

    Moreau P, Robillard N, Jego G, Pellat C, Le Gouill S, Thoumi S et al (2006) Lack of CD27 in myeloma delineates different presentation and outcome. Br J Haematol 2:168–170

    Article  Google Scholar 

  35. 35.

    Cannizzo E, Bellio E, Sohani AR, Hasserjian RP, Ferry JA, Dorn ME et al (2010) Multiparameter immunophenotyping by flow cytometry in multiple myeloma: The diagnostic utility of defining ranges of normal antigenic expression in comparison to histology. Cytom B Clin Cytom 4:231–238

    Google Scholar 

  36. 36.

    Tohami T, Drucker L, Shapiro H, Radnay J, Lishner M (2007) Overexpression of tetraspanins affects multiple myeloma cell survival and invasive potential. FASEB J 3:691–699

    Article  CAS  Google Scholar 

  37. 37.

    Barrena S, Almeida J, Yunta M, Lopez A, Fernandez-Mosteirin N, Giralt M et al (2005) Aberrant expression of tetraspanin molecules in B-cell chronic lymphoproliferative disorders and its correlation with normal B-cell maturation. Leukemia 8:1376–1383

    Article  CAS  Google Scholar 

  38. 38.

    Luo RF, Zhao S, Tibshirani R, Myklebust JH, Sanyal M, Fernandez R et al (2010) CD81 protein is expressed at high levels in normal germinal center B cells and in subtypes of human lymphomas. Hum Pathol 2:271–280

    Article  CAS  Google Scholar 

  39. 39.

    Tembhare PR, Yuan CM, Venzon D, Braylan R, Korde N, Manasanch E et al (2014) Flow cytometric differentiation of abnormal and normal plasma cells in the bone marrow in patients with multiple myeloma and its precursor diseases. Leuk Res 3:371–376

    Article  Google Scholar 

  40. 40.

    Alapat D, Coviello-Malle J, Owens R, Qu P, Barlogie B, Shaughnessy JD et al (2012) Diagnostic usefulness and prognostic impact of CD200 expression in lymphoid malignancies and plasma cell myeloma. Am J Clin Pathol 1:93–100

    Article  Google Scholar 

  41. 41.

    Olteanu H, Harrington AM, Kroft SH (2012) CD200 expression in plasma cells of nonmyeloma immunoproliferative disorders: clinicopathologic features and comparison with plasma cell myeloma. Am J Clin Pathol 6:867–876

    Article  CAS  Google Scholar 

  42. 42.

    Moreaux J, Hose D, Reme T, Jourdan E, Hundemer M, Legouffe E et al (2006) CD200 is a new prognostic factor in multiple myeloma. Blood 13:4194–4197

    Article  CAS  Google Scholar 

Download references

Author information

Affiliations

Authors

Corresponding author

Correspondence to Dominik F. Draxler.

Ethics declarations

Conflict of interest

The authors declare they have no conflict of interest with respect to the present manuscript.

Ethical Approval

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

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary material 1 (DOCX 21 kb)

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Draxler, D.F., Wutzlhofer, L.M., Slavka, G. et al. Flow Cytometric Evaluation of Traditional and Novel Surface Markers for the Diagnosis of Plasma Cell Dyscrasias. Indian J Hematol Blood Transfus 35, 673–682 (2019). https://doi.org/10.1007/s12288-019-01105-w

Download citation

Keywords

  • Multiple myeloma
  • Plasma cell dyscrasias
  • Plasma cells
  • FACS
  • Flow cytometry