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Immunophenotyping of Plasma Cells in Multiple Myeloma

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Multiple Myeloma

Abstract

Multiparametric immunophenotyping of multiple myeloma (MM) and other plasma cell (PC) dyscrasias represents an attractive approach not only for research purposes but also in clinical practice. Based on well-established antigenic patterns, discrimination between myelomatous and normal PCs can be easily achieved in various types of samples, and this can be particularly valuable for the differential diagnosis between MGUS and MM and for monitoring residual disease in the latter. In addition, immunophenotyping may be an alternative and more reproducible method than morphology for evaluating PC infiltration, as well as for specifically analyzing DNA content and the cell-cycle distribution of different subsets of PCs. Despite the widespread use, standardization of methods and protocols still remains a challenge. In this chapter, we describe in detail the protocols and precise instructions for specimen collection, sample preparation, together with the methods for staining PCs and flow cytometry, data acqui- sition, and data analysis, including the more recent developments in the field. We highlight the most frequent limitations, and provide troubleshooting and practical recommendations that could help to solve them. The goal of this chapter is to emphasize the relevance of methodological issues in order to obtain reproducible and high-quality results regarding the phenotypic analysis of PCs.

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References

  1. Harada H., Kawano M. M., Huang N., et al. (1993) Phenotypic difference of normal plasma cells from mature myeloma cells. Blood 81, 2658–2663.

    PubMed  CAS  Google Scholar 

  2. Terstappen L. W. (1990) Identification and characterization of plasma cells in normal human bone marrow by high-resolution flow cytometry. Blood 76, 1739–1747.

    PubMed  CAS  Google Scholar 

  3. San Miguel J. F. (1991) Immunophenotypic heterogeneity of multiple myeloma: influence on the biology and clinical course of the disease. Castellano-Leones (Spain) Cooperative Group for the Study of Monoclonal Gammopathies. Br. J. Haematol. 77, 185–190.

    Article  PubMed  CAS  Google Scholar 

  4. Ocqueteau M., Orfao A., Almeida J., 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. 152, 1655–1665.

    PubMed  CAS  Google Scholar 

  5. Almeida J., Orfao A., Mateo G., et al. (1999) Immunophenotypic and DNA content characteristics of plasma cells in multiple myeloma and monoclonal gammopathy of undetermined significance. Pathol. Biol. (Paris) 47, 119–127.

    CAS  Google Scholar 

  6. Rawstron A. C., Davies F. E., DasGupta R., et al. (2002) Flow cytometric disease monitoring in multiple myeloma: the relationship between normal and neoplastic plasma cells predicts outcome after transplantation. Blood 100, 3095–3100.

    Article  PubMed  CAS  Google Scholar 

  7. San Miguel J. F., Almeida J., Mateo G., et al. (2002) Immunophenotypic evaluation of the plasma cell compartment in multiple myeloma: a tool for comparing the efficacy of different treatment strategies and predicting outcome. Blood 99, 1853–1856.

    Article  PubMed  Google Scholar 

  8. Almeida J., Orfao A., Ocqueteau M., et al. (1999) High-sensitive immunophenotyping and DNA ploidy studies for the investigation of minimal residual disease in multiple myeloma. Br. J. Haematol. 107, 121–131.

    Article  PubMed  CAS  Google Scholar 

  9. Pope B., Brown R., Gibson J., and Joshua D. (1997) Plasma cells in peripheral blood stem cell harvests from patients with multiple myeloma are predominantly polyclonal. Bone Marrow Transplant. 20, 205–210.

    Article  PubMed  CAS  Google Scholar 

  10. Mateo G., Corral M., Almeida J., et al. (2003) Immunophenotypic analysis of peripheral blood stem cell harvests from patients with multiple myeloma. Haematologica 88, 1013–1021.

    PubMed  Google Scholar 

  11. Rawstron A. C., Barrans S. L., Blythe D., et al. (2001) In multiple myeloma, only a single stage of neoplastic plasma cell differentiation can be identified by VLA-5 and CD45 expression. Br. J. Haematol. 113, 794–802.

    Article  PubMed  CAS  Google Scholar 

  12. Jego G., Robillard N., Puthier D., et al. (1999) Reactive plasmacytoses are expansions of plasmablasts retaining the capacity to differentiate into plasma cells. Blood 94, 701–712.

    PubMed  CAS  Google Scholar 

  13. Garcia-Sanz R., Orfao A., Gonzalez M., et al. (1999) Primary plasma cell leukemia: clinical, immunophenotypic, DNA ploidy, and cytogenetic characteristics. Blood 93, 1032–1037.

    PubMed  CAS  Google Scholar 

  14. Mason D., Andre P., Bensussan A., et al. (2002) CD antigens 2002. Blood 99, 3877–3880.

    Article  PubMed  CAS  Google Scholar 

  15. Papa S. (2002) European Working Group on Clinical Cell Analysis (EWGCCA): 6 years on. J. Biol. Regul. Homeost. Agents 16, 253–256.

    PubMed  CAS  Google Scholar 

  16. Kraan J., Gratama J. W., Keeney M., and D’Hautcourt J. L. (2003) Setting up and calibration of a flow cytometer for multicolor immunophenotyping. J. Biol. Regul. Homeost. Agents 17, 223–233.

    PubMed  CAS  Google Scholar 

  17. Dahl I. M., Rasmussen T., Kauric G., and Husebekk A. (2002) Differential expression of CD56 and CD44 in the evolution of extramedullary myeloma. Br. J. Haematol. 116, 273–277.

    Article  PubMed  CAS  Google Scholar 

  18. San Miguel J. E., Vidriales M. B., Ocio E., et al. (2003) Immunophenotypic analysis of Waldenstrom’s macroglobulinemia. Semin. Oncol. 30, 187–195.

    Article  PubMed  CAS  Google Scholar 

  19. Radbruch A. (2000) Flow Cytometry and Cell Sorting, Springer Verlag, New York.

    Google Scholar 

  20. Drexler H. G. and Matsuo Y. (2000) Malignant hematopoietic cell lines: in vitro models for the study of multiple myeloma and plasma cell leukemia. Leuka. Res. 24, 681–703.

    Article  CAS  Google Scholar 

  21. Jourdan M., Ferlin M., Legouffe E., et al. (1998) The myeloma cell antigen syndecan-1 is lost by apoptotic myeloma cells. Br. J. Haematol. 100, 637–646.

    Article  PubMed  CAS  Google Scholar 

  22. Bharti A. C., Shishodia S., Reuben J. M., et al. (2003) Nuclear factor-κB and STAT3 are constitutively active in CD138+ cells derived from multiple myeloma patients, and their suppression leads to apoptosis. Blood 103, 3175–3184.

    Article  PubMed  Google Scholar 

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Manzanera, G.M., San Miguel Izquierdo, J.F., de Matos, A.O. (2005). Immunophenotyping of Plasma Cells in Multiple Myeloma. In: Brown, R.D., Ho, P.J. (eds) Multiple Myeloma. Methods in Molecular Medicine™, vol 113. Humana Press. https://doi.org/10.1385/1-59259-916-8:5

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  • DOI: https://doi.org/10.1385/1-59259-916-8:5

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-392-3

  • Online ISBN: 978-1-59259-916-5

  • eBook Packages: Springer Protocols

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