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Original Article

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Korean J Hematol 2012; 47(4):

Published online December 31, 2012

https://doi.org/10.5045/kjh.2012.47.4.260

© The Korean Society of Hematology

Simplified flow cytometric immunophenotyping panel for multiple myeloma, CD56/CD19/CD138(CD38)/CD45, to differentiate neoplastic myeloma cells from reactive plasma cells

Tae-Dong Jeong1, Chan-Jeoung Park1*, Hyoeun Shim2, Seongsoo Jang1, Hyun-Sook Chi1, Dok Hyun Yoon3, Dae-Young Kim3, Jung-Hee Lee3, Je-Hwan Lee3, Cheolwon Suh3, and Kyoo Hyung Lee3

Author Info. +

1Department of Laboratory Medicine, University of Ulsan College of Medicine and Asan Medical Center, Seoul, Korea.

2Department of Laboratory Medicine, Ajou University School of Medicine, Suwon, Korea.

3Department of Internal Medicine, University of Ulsan College of Medicine and Asan Medical Center, Seoul, Korea.

Correspondence to : Correspondence to Chan-Jeoung Park, M.D., Ph.D. Department of Laboratory Medicine, University of Ulsan College of Medicine and Asan Medical Center, 88, Olympic-ro 43-gil, Songpa-gu, Seoul 138-736, Korea. Tel: +82-2-3010-4508, Fax: +82-2-478-0884, cjpark@amc.seoul.kr

Received: July 16, 2012; Revised: September 27, 2012; Accepted: November 6, 2012

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

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Background

Flow cytometric immunophenotyping has been used to identify neoplastic plasma cell populations in patients with multiple myeloma (MM). Previous reports have described the use of several antigens, including CD38, CD138, CD56, CD117, CD52, CD19 and CD45, to distinguish distinct populations of plasma cells. The aim of this study was to evaluate a simplified immunophenotyping panel for MM analysis.

Methods

A total of 70 patients were enrolled in the study, 62 of which were newly diagnosed with MM (untreated), whereas the remaining 8 were undergoing bone marrow assessment as part of follow-up after treatment (treated). Treated cases included 3 patients with relapse and 5 patients with persistence of MM. Multiparametric flow cytometric immunophenotyping was performed using monoclonal antibodies against CD56, CD19, CD138 (CD38), and CD45.

Results

In differential counts, plasma cells in bone marrow (BM) accounted for 3.6-93.2% of the total nucleated cell count. The positive expression rates of CD56, CD19, CD138, and CD45 in neoplastic myeloma cells were 83.9%, 0%, 98.4%, and 37.1%, respectively, among the 62 untreated cases, and 75.0%, 0%, 87.5%, and 37.5%, respectively, among the 8 treated cases. CD19 expression of neoplastic plasma cells was negative in both untreated and treated cases.

Conclusion

The simplified immunophenotyping panel, CD56/CD19/CD138(CD38)/CD45, is useful for distinguishing neoplastic myeloma cells from reactive plasma cells in clinical practice. In addition, CD19 represents the most valuable antigen for identifying neoplastic myeloma cells in patients with MM.

Keywords Multiple myeloma, Flow cytometry, Immunophenotyping, Neoplastic plasma cells, CD19 negativity

Article

Original Article

Korean J Hematol 2012; 47(4): 260-266

Published online December 31, 2012 https://doi.org/10.5045/kjh.2012.47.4.260

Copyright © The Korean Society of Hematology.

Simplified flow cytometric immunophenotyping panel for multiple myeloma, CD56/CD19/CD138(CD38)/CD45, to differentiate neoplastic myeloma cells from reactive plasma cells

Tae-Dong Jeong1, Chan-Jeoung Park1*, Hyoeun Shim2, Seongsoo Jang1, Hyun-Sook Chi1, Dok Hyun Yoon3, Dae-Young Kim3, Jung-Hee Lee3, Je-Hwan Lee3, Cheolwon Suh3, and Kyoo Hyung Lee3

1Department of Laboratory Medicine, University of Ulsan College of Medicine and Asan Medical Center, Seoul, Korea.

2Department of Laboratory Medicine, Ajou University School of Medicine, Suwon, Korea.

3Department of Internal Medicine, University of Ulsan College of Medicine and Asan Medical Center, Seoul, Korea.

Correspondence to: Correspondence to Chan-Jeoung Park, M.D., Ph.D. Department of Laboratory Medicine, University of Ulsan College of Medicine and Asan Medical Center, 88, Olympic-ro 43-gil, Songpa-gu, Seoul 138-736, Korea. Tel: +82-2-3010-4508, Fax: +82-2-478-0884, cjpark@amc.seoul.kr

Received: July 16, 2012; Revised: September 27, 2012; Accepted: November 6, 2012

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Background

Flow cytometric immunophenotyping has been used to identify neoplastic plasma cell populations in patients with multiple myeloma (MM). Previous reports have described the use of several antigens, including CD38, CD138, CD56, CD117, CD52, CD19 and CD45, to distinguish distinct populations of plasma cells. The aim of this study was to evaluate a simplified immunophenotyping panel for MM analysis.

Methods

A total of 70 patients were enrolled in the study, 62 of which were newly diagnosed with MM (untreated), whereas the remaining 8 were undergoing bone marrow assessment as part of follow-up after treatment (treated). Treated cases included 3 patients with relapse and 5 patients with persistence of MM. Multiparametric flow cytometric immunophenotyping was performed using monoclonal antibodies against CD56, CD19, CD138 (CD38), and CD45.

Results

In differential counts, plasma cells in bone marrow (BM) accounted for 3.6-93.2% of the total nucleated cell count. The positive expression rates of CD56, CD19, CD138, and CD45 in neoplastic myeloma cells were 83.9%, 0%, 98.4%, and 37.1%, respectively, among the 62 untreated cases, and 75.0%, 0%, 87.5%, and 37.5%, respectively, among the 8 treated cases. CD19 expression of neoplastic plasma cells was negative in both untreated and treated cases.

Conclusion

The simplified immunophenotyping panel, CD56/CD19/CD138(CD38)/CD45, is useful for distinguishing neoplastic myeloma cells from reactive plasma cells in clinical practice. In addition, CD19 represents the most valuable antigen for identifying neoplastic myeloma cells in patients with MM.

Keywords: Multiple myeloma, Flow cytometry, Immunophenotyping, Neoplastic plasma cells, CD19 negativity

Fig 1.

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Figure 1.

Typical laboratory protocol for distinguishing neoplastic plasma cells (CD56+or-/CD19-/CD138+/CD45-) from reactive plasma cells (CD56-/CD19+/CD138+/CD45+) in patients with multiple myeloma. Plasma cells were gated with CD138+ and low side scatter and were distinguished as neoplastic plasma cells (CD56+/CD19-/CD138+/CD45-) and reactive plasma cells (CD56-/CD19+/CD138+/CD45+).

Blood Research 2012; 47: 260-266https://doi.org/10.5045/kjh.2012.47.4.260

Fig 2.

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Figure 2.

Flow cytometric immunophenotyping of 2 cases (A and B) with CD138-negative neoplastic plasma cells. Plasma cells were gated with CD38+ and low side scatter, followed by selection of CD45-dim (A) and CD45-negative (B) cells. In the both cases (A and B), neoplastic plasma cells showed CD19- and CD56+ expression (A, dim and B, intermediate).

Blood Research 2012; 47: 260-266https://doi.org/10.5045/kjh.2012.47.4.260

Fig 3.

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Figure 3.

Comparison of the monoclonal immunoglobulin component (M-component) and the tumor burden of neoplastic plasma cells (PCs) in bone marrow (BM) aspirates according to flow cytometry (FCM). (A) A weak positive correlation (r=0.15) was noted between the fraction of neoplastic PCs according to FCM and the M-component, but this was not statistical significant (P=0.245). (B) The proportion of neoplastic PCs in BM aspirates, i.e., fraction of neoplastic PCs according to FCM×percentage of PCs among the total number of nucleated cells in the BM aspirates according to differential cell count, showed a statistically significant positive correlation with the M-component (r=0.26, P=0.045).

Blood Research 2012; 47: 260-266https://doi.org/10.5045/kjh.2012.47.4.260

Table 1 . Patient demographics..

a)Continuous variables were analyzed using the Mann-Whitney U Test, and categorical variables were analyzed using the Fisher's exact test..

b)Differential cell count of BM aspirates..

Abbreviations: CRP, C-reactive protein; Hb, hemoglobin; Ig, immunoglobulin; ISS, international staging system; LD, lactate dehydrogenase; NS, not significant..

Table 2 . Comparison of overall positive expression rates for CD56, CD19, CD138, and CD45 in neoplastic myeloma cells between the untreated and treated patients..

Abbreviations: NA, not available; Neg, negative; Pos, positive..

Table 3 . Clinical and laboratory characteristics of 2 cases with CD138-negative neoplastic plasma cells according to FCM immunophenotyping..

Abbreviations: EP, electrophoresis; IFE, immunofixation electrophoresis..

Table 4 . Comparison of the proportion of neoplastic plasma cells between the untreated and treated patients..


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