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Updated: Aug 17, 2026
Flow Cytometric Characterization of Murine B Cell Development
Published on: January 22, 2021
Demonstration of a germinal center immunophenotype in lymphomas by immunocytochemistry and flow cytometry
Andrew Jack1, Sharon Barrans, David Blythe
1Leeds Teaching Hospitals NHS Trust, UK.
Insights
Most B-cell lymphomas originate from the germinal center. Immunocytochemistry and flow cytometry can distinguish germinal center B-cell lymphomas from post-germinal center types, aiding in prognosis and classification.
Area of Science:
- Immunology
- Hematology
- Oncology
Background:
- The germinal center is crucial in B-cell lymphoma pathogenesis.
- Most B-cell lymphomas are derived from germinal center or post-germinal center stages.
- Germinal center-type diffuse large B-cell lymphomas show a better prognosis.
Purpose of the Study:
- To differentiate B-cell lymphomas based on germinal center origin.
- To evaluate immunophenotypic methods for routine diagnostics.
- To improve classification of mature B-cell malignancies.
Main Methods:
- Utilized immunocytochemistry for morphological identification.
- Employed flow cytometry for precise cell typing with multiple antibodies.
- Compared antibody-based methods with microarray technology.
Main Results:
- B-cell lymphomas can be divided into germinal center and post-germinal center phenotypes.
- Immunocytochemistry and flow cytometry are practical diagnostic tools.
- These methods offer simpler and cheaper alternatives to microarrays.
Conclusions:
- Immunophenotypic methods can classify B-cell lymphomas.
- Distinguishing germinal center phenotypes aids in prognosis.
- Further understanding of germinal center biology will refine classification.
Abstract:
The germinal center plays an important role in the pathogenesis of B-cell lymphomas, and evidence exists to suggest that most cases are germinal center or postgerminal center derived. Burkitt lymphoma and follicular lymphoma are derived from the germinal center stage of differentiation. It has been shown that diffuse large B-cell lymphomas with a germinal center-type pattern of RNA expression or a germinal center cell phenotype using immunocytochemistry have a more favorable outcome compared with those with a postgerminal center/activated profile. Microarray technology may not be available in many diagnostic laboratories, and antibody-based methods are much simpler and cheaper and are therefore more applicable to the routine setting. Immunocytochemistry has the advantage that the cells of interest are identified morphologically, and it is applicable retrospectively to fixed tissue. The main disadvantage is that only single-color staining is currently used in the routine setting. Flow cytometry allows one to obtain a more precise definition of individual cell types. The cells of interest are identified by a combination of physical characteristics and by the use of multiple antibodies labeled with different fluorochromes. Flow cytometry has the major advantage of being able to analyze very large numbers of cells, and results can be obtained within a few hours of the specimen being taken. The methods described allow B-cell lymphomas to be crudely divided into two groups, those with a germinal center phenotype and those that are mainly postgerminal center tumors. As knowledge of the normal biology of the germinal center develops, it will become possible to use immunophenotypic methods to more precisely classify all types of mature B-cell malignancies.
