Korean J Hematol.  2012 Dec;47(4):260-266. 10.5045/kjh.2012.47.4.260.

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

• Jeong TD
• Park CJ
• Shim H
• Jang S
• Chi HS
• Yoon DH
• Kim DY
• Lee JH
• Lee JH
• Suh C
• Lee KH

• KMID: 1832104
• DOI: http://doi.org/10.5045/kjh.2012.47.4.260

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.

Keyword

Multiple myeloma; Flow cytometry; Immunophenotyping; Neoplastic plasma cells; CD19 negativity

MeSH Terms

Antibodies, Monoclonal
Bone Marrow
Cell Count
Flow Cytometry
Follow-Up Studies
Humans
Immunophenotyping
Multiple Myeloma
Plasma
Plasma Cells
Recurrence
Antibodies, Monoclonal

Figure

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  Fig. 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+).

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  Fig. 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).

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  Fig. 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).

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