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Published on: October 19, 2014
Molecular characterization of chronic lymphocytic leukemia with two distinct cell populations: Evidence for separate
1Department of Laboratory Hematology, University Health Network and University of Toronto, Toronto, Canada.
Insights
This study identified two distinct B-cell clones in a patient with persistent lymphocytosis. Flow cytometry and genetic analysis confirmed separate origins and unique characteristics for each clone.
Area of Science:
- Hematology
- Immunology
- Genetics
Background:
- Persistent lymphocytosis can indicate underlying hematologic disorders.
- Flow cytometric immunophenotyping is crucial for characterizing lymphocyte populations.
- Distinguishing distinct B-cell clones is important for accurate diagnosis and management.
Observation:
- An elderly woman presented with persistent lymphocytosis.
- Flow cytometry revealed two distinct B-cell populations with differing kappa and lambda light chain expression.
- One clone showed CD19 dim, CD5 bright, CD23 moderate, kappa+; the other showed CD19 moderate, CD5 bright, CD23 dim, lambda+.
Findings:
- Two separate B-cell clones were isolated using flow cytometry.
- Polymerase chain reaction (PCR) analysis of immunoglobulin heavy chain (IgH) genes showed distinct rearrangements for each clone.
- Fluorescence in situ hybridization (FISH) identified trisomy 12 in the lambda-restricted clone, while the kappa-restricted clone had normal FISH patterns.
Implications:
- This case highlights the utility of multi-parameter analysis in identifying distinct B-cell clones.
- The findings underscore the importance of combining immunophenotyping with molecular and cytogenetic techniques.
- Understanding clonal heterogeneity is vital for diagnosing and potentially treating lymphoproliferative disorders.
Abstract:
We report a case of an elderly woman with persistent lymphocytosis in whom flow cytometric immunophenotyping revealed 2 distinct clonal B-cell populations with different light chain restrictions. One clone was CD19+ (dim), CD5+ (bright), CD23+ (moderate), and kappa+; the other clone was CD19+ (moderate), CD5+ (bright), CD23+ (dim), and lambda+. We separated the 2 clones by flow cytometry from peripheral blood lymphocytes for further molecular and cytogenetic analysis. Polymerase chain reaction (PCR) analysis of the immunoglobulin heavy chain (IgH) genes revealed that the clones had bands of slightly different size. The PCR products of the framework 3 (FR3) region were cloned, and sequence analysis confirmed that each population had distinct, clone-specific IgH gene rearrangements. Fluorescence in situ hybridization (FISH) analysis revealed trisomy 12 in the lambda-restricted B-cell clone, whereas the k-restricted population had normal FISH patterns. Our results demonstrated that the immunophenotypically different cell populations originated from 2 separate clones.
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