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Updated: Sep 6, 2025
Flow Cytometric Characterization of Murine B Cell Development
Published on: January 22, 2021
Immunophenotypic Characterization and Ploidy Analysis of Neoplastic Plasma Cells by Multiparametric Flow Cytometry
1Department of Hematology, Sanjay Gandhi Post Graduate Institute of Medical Sciences (SGPGIMS), Lucknow, Uttar Pradesh India.
Insights
Flow cytometry immunophenotyping and ploidy analysis of plasma cells in myeloma patients reveal aberrant antigen expression and hyperdiploidy in most cases. CD81 expression correlates with disease burden, suggesting its prognostic value.
Area of Science:
- Hematology
- Immunology
- Oncology
Background:
- Flow cytometric immunophenotyping is crucial for diagnosing and prognosticating plasma cell dyscrasias.
- Understanding plasma cell immunophenotype and ploidy status is vital for myeloma management.
Purpose of the Study:
- To evaluate the immunophenotype and ploidy status of plasma cells in myeloma patients.
- To correlate these findings with laboratory parameters and assess prognostic potential.
Main Methods:
- Bone marrow samples from 70 newly diagnosed myeloma patients were analyzed using flow cytometry.
- A panel of antibodies including CD138, CD38, CD19, CD45, CD28, CD81, CD56, CD200, and CD229 was employed.
- FxCycle Violet dye was used for ploidy analysis of clonal plasma cells.
Main Results:
- A positive correlation was found between morphological and FCM-based plasma cell enumeration.
- Aberrant expression of CD56 (88.5%), CD200 (77%), and CD81 (23%) was frequently observed.
- Hyperdiploid plasma cells were identified in 80% of cases; CD81 expression correlated with higher monoclonal protein and renal insufficiency.
- CD229 was expressed in all cases and is a reliable gating marker; CD56/CD200 and CD45- can identify abnormal plasma cells.
Conclusions:
- Flow cytometry immunophenotyping and ploidy analysis provide valuable diagnostic and prognostic information in myeloma.
- CD81 may serve as a prognostic marker, correlating with disease burden.
- Ploidy analysis can aid in identifying patients with poor prognosis and should be considered in routine workup.
Abstract:
Flow cytometric (FCM) immunophenotyping is an important tool for generating diagnostic and prognostic information in plasma cell dyscrasias. This study aimed to evaluate the immunophenotype and ploidy status of plasma cells (PCs) in patients of myeloma and its correlation with other laboratory parameters. Bone marrow of 70 newly diagnosed cases of myeloma were subjected to FCM using a panel of antibodies; CD138, CD38, CD19, CD45, CD28, CD81, CD56, CD200, and CD229. FxCycle Violet (FCV) dye was used for the ploidy analysis of clonal PCs. Median age was 60 years with M:F ratio of 3.2:1. A positive correlation was noted between the morphological and FCM-based PC enumeration (r = 0.4, p = 0.001). Aberrant expression of CD56, CD200, CD28, CD117, CD81 and CD19 and was observed in 88.5%, 77%, 29%, 37%, 23% and 17% cases respectively. Two aberrant antigens were noted in all cases. CD81 + cases had a relatively higher quantity of monoclonal-protein (> 1 g/dl, p < 0.05) and renal insufficiency (Cr > 2 mg/dl, p < 0.05) as compared to the CD81- cases. CD229 was expressed in all the cases, with a median MFI in PCs significantly higher than other hematopoietic elements. Hyperdiploid PCs (median DI-1.59, range, 1.16-2.6) were noted in 80% cases (n = 48), diploid/ near-hyperdiploid PCs in 8% (n = 5) cases and hypodiploidy in 3% (n = 1) cases. Bright CD56/CD200 and CD45- can identify abnormal PC in the majority of the cases. CD81 appears to correlate with disease burden and might be useful as a prognostic marker. CD229 is a reliable gating marker for plasma cells. Ploidy analysis may be incorporated in routine workup to guide in the identification of patients with poor prognosis.
Supplementary Information:
The online version contains supplementary material available at 10.1007/s12288-021-01477-y.
