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Immunophenotyping large B-cell lymphomas. Flow cytometric pitfalls and pathologic correlation
H C Bertram1, I J Check, M A Milano
1Department of Pathology, Evanston Northwestern Healthcare, Northwestern University Medical School, Evanston, IL, USA.
Insights
Diagnosing large cell lymphomas using flow cytometry (FC) can be challenging. This study found that morphologic features do not predict diagnostic difficulty, emphasizing critical gating strategies for improved FC yield in lymphoma diagnosis.
Area of Science:
- Hematology
- Oncology
- Immunology
Background:
- Large cell lymphomas present diagnostic challenges for flow cytometry (FC).
- Accurate FC protocols are crucial for effective lymphoma diagnosis and classification.
- Correlating FC data with histological features aids in understanding diagnostic complexities.
Purpose of the Study:
- To refine diagnostic protocols for large cell lymphomas using FC.
- To establish correlations between FC findings and demographic/histologic features.
- To enhance the diagnostic accuracy of FC in identifying large cell lymphomas.
Main Methods:
- Reviewed 63 cases of large B-cell lymphoma diagnosed between 1995 and 1999.
- Analyzed diagnostic slides, FC light scatter, and staining patterns.
- Categorized lymphomas into "clear cut" and "complex" light scatter patterns.
Main Results:
- 51 lymphomas had adequate material for review, with 33% in the "clear cut" pattern and 67% in the "complex" pattern.
- "Clear cut" cases showed higher mitotic activity and cellularity.
- Histological features like apoptosis, necrosis, and sclerosis were common, irrespective of FC patterns.
Conclusions:
- Morphologic features of large cell lymphomas do not reliably predict diagnostic challenges in FC.
- Effective gating strategies are essential for improving diagnostic yield in FC analysis of lymphomas.
- FC remains a valuable tool, but careful protocol optimization is necessary for complex cases.
Abstract:
Large cell lymphomas often challenge the diagnostic flow cytometrist. The purposes of this study were to improve our protocols for diagnosing large cell lymphomas and to correlate flow cytometric (FC) data with demographic and histologic features. We identified 63 cases of large B-cell lymphoma between January 1, 1995, and July 30, 1999, and reviewed the diagnostic slides and FC light scatter and staining patterns. The 51 lymphomas with adequate material for systemic review fell into 2 light scatter patterns: "clear cut," with large abnormal cells (high forward scatter relative to normal lymphocytes), 17 cases (33%); and "complex," 34 cases (67%). Clear-cut cases were more mitotically active (average of 42 vs 25 per 10 high-power fields), with higher cellularity. Apoptosis, geographic necrosis, and sclerosis were present histologically in many cases, regardless of FC findings. We conclude that morphologic features of large cell lymphomas do not predict which cases will be difficult to diagnose by FC. Gating strategies can be critical to improve the diagnostic yield.
