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Updated: May 7, 2026
Flow Cytometric Characterization of Murine B Cell Development
Published on: January 22, 2021
Multiparameter flow cytometry is necessary for detection, characterization and diagnostics of composite mature B-cell
Sanja Perković1, Sandra Bašić-Kinda, Igor Aurer
1Clinical Unit for Cellular Immunodiagnostics and In Vitro Procedures, Department of Laboratory Diagnostics, University Hospital Centre Zagreb, School of Medicine, University of Zagreb, Kišpatićeva 12, 10000, Zagreb, Croatia, sanja.perkovic@gmail.com.
Insights
Composite mature B-cell lymphoproliferative neoplasms are rare. This study found biclonality in 3.9% of patients, particularly in hairy cell leukemia cases, challenging traditional diagnostic methods.
Area of Science:
- Hematology
- Oncology
- Immunophenotyping
Background:
- Composite mature B-cell lymphoproliferative neoplasms involve multiple distinct B-cell malignancies.
- Accurate diagnosis is crucial for effective treatment and patient outcomes.
Purpose of the Study:
- To immunophenotypically profile composite mature B-cell lymphoproliferative neoplasms.
- To evaluate the frequency and characteristics of biclonality in these disorders.
- To identify reliable diagnostic markers for biclonal lymphomas.
Main Methods:
- Retrospective analysis of 413 patients with de novo leukemic B-cell chronic lymphoproliferative disorders.
- Multiparametric flow cytometry for immunophenotypic profiling.
- Analysis of aberrant B-cell phenotypes and light chain restriction.
Main Results:
- Biclonality was identified in 3.9% of patients.
- Hairy cell leukemia cases showed a significantly higher frequency of biclonality (22%).
- Traditional kappa/lambda light chain ratio was unreliable for detecting biclonality.
Conclusions:
- Composite lymphomas require advanced diagnostic techniques beyond the kappa/lambda ratio.
- Aberrant B-cell phenotypes and specific cell surface antigens (CD22, CD20, IgM, CD23) are key for discrimination.
- Multiparametric flow cytometry is essential for identifying coexisting malignant clones.
Abstract:
Composite mature B-cell lymphoproliferative neoplasms are rare entities characterized by the simultaneous presence of two or more distinctive B-cell derived monoclonal malignancies. This retrospective study used multiparametric flow cytometric analysis aimed at immunophenotypic profiling of composite mature B-cell lymphoproliferative neoplasms in a cohort of 413 subsequent patients with de novo leukemic B-cell chronic lymphoproliferative disorders diagnosed in our institution during a 30-month period. Biclonality was found in 16 (3.9 %) patients. The vast majority (88 %) of the cases had one of the clones phenotypically corresponding to chronic lymphocytic leukemia/small lymphocytic lymphoma (CLL/SLL). Only when composite cases were categorized by phenotype of the non-CLL/SLL malignant population did we find a statistically significant (P = 0.001) higher frequency of biclonality among cases with hairy cell leukemia (22 %). Biclonal cases had the overall B-cell membrane κ to λ ratio within the normal range (median, 1.9; reference interval 0.5-4.0), making recognition of malignancy somewhat challenging. Our analysis strategy was therefore based on the detection of aberrant B-cell phenotypes, with subsequent confirmation of the monoclonal nature of neoplastic clones with regards to light chain restriction analysis. Discrimination of the coexisting clones in biclonal cases was possible on the basis of the expression of other antigen(s) (63 %), light scatter properties (44 %), different surface light chain restriction (69 %) and/or pattern of expression (44 %). The most informative cell surface antigens proved to be CD22, CD20, surface IgM, and CD23. In conclusion, historic κ/λ ratio is not a reliable approach and is a poor measurement for the detection of composite lymphomas. More creative analysis techniques should be utilized for this purpose.
