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Updated: Aug 12, 2026
Detection and Enrichment of Rare Antigen-specific B Cells for Analysis of Phenotype and Function
Published on: February 16, 2017
Detection of immunophenotypic abnormalities in paraffin-embedded B-lineage non-Hodgkin's lymphomas
A B Gelb1, R V Rouse, R F Dorfman
1Department of Pathology, Stanford University School of Medicine, California.
Insights
This study details immunophenotypic abnormalities in B-lineage non-Hodgkin
Area of Science:
- Hematopathology
- Immunohistochemistry
- Oncology
Background:
- Accurate diagnosis of B-lineage non-Hodgkin's lymphomas (NHL) relies on identifying specific immunophenotypic markers.
- Routine paraffin-embedded tissues present challenges for immunophenotyping, necessitating optimized antibody panels.
- Understanding the frequency and patterns of immunophenotypic abnormalities aids in subtyping and diagnosing NHL.
Purpose of the Study:
- To analyze the frequency of immunophenotypic abnormalities in a large cohort of paraffin-embedded B-lineage NHL.
- To evaluate the utility of specific markers (CD20, CD45RA, Ig light chain, CD43, CD45RO, CD3, bcl-2) in diagnosing NHL subtypes.
- To assess the diagnostic value of combining multiple immunophenotypic criteria.
Main Methods:
- Immunohistochemical analysis of 1,474 routinely fixed, paraffin-embedded B-lineage NHL cases.
- Determination of B-lineage using CD20, CD45RA, and immunoglobulin (Ig) light chain restriction.
- Assessment of CD43 coexpression, Ig light chain restriction, and bcl-2 overexpression in various NHL subtypes.
Main Results:
- CD43 coexpression was the most common abnormality (e.g., 60% in mantle cell lymphomas).
- Ig light chain restriction was frequent in specific subtypes (e.g., 67% in plasmacytoid diffuse small cell lymphomas).
- bcl-2 overexpression was observed in 71% of follicular lymphomas but not in reactive hyperplasias.
Conclusions:
- Immunophenotypic analysis of paraffin-embedded tissues is crucial for diagnosing B-lineage NHL.
- Specific markers like CD43 and Ig light chain restriction are valuable for identifying NHL subtypes.
- Combining diagnostic criteria enhances the sensitivity of immunodiagnosis for NHL.
Abstract:
The authors analyzed the frequency of immunophenotypic abnormalities in 1,474 cases of routinely fixed, paraffin-embedded B-lineage non-Hodgkin's lymphomas. B-lineage was determined by immunoreactivity for CD20 (L26, 92%); CD45RA (4KB5, an additional 3%) or immunoglobulin (Ig) light chain restriction (remaining 5%). CD45RA was found to be especially helpful on Bouin's-fixed or decalcified tissue and Ig staining was most helpful in plasmacytoid lesions. Coexpression of the T-cell marker CD43 (Leu-22) was the most common immunophenotypic abnormality, seen in 60% of mantle cell lymphomas (MCL), 39% of CLL/small lymphocytic lymphomas, 16% of diffuse large cell lymphomas (DLCL), but only 5% of follicular lymphomas (FL). Antibodies to CD45RO (A6 and UCHL1) and CD3 (polyclonal) were useful in distinguishing infiltrating T cells from B cells coexpressing CD43. Ig light chain restriction was the next commonest immunophenotypic abnormality, which was identified in 67% of plasmacytoid diffuse small cell lymphomas, 43% of MCLs, 35% of monocytoid B-cell lymphomas and 28% of FLs. Overexpression of bcl-2 oncogenic protein was observed in 71% of FLs (n = 96), but not in a control group of reactive follicular hyperplasias (n = 34). Combining two criteria increased the sensitivity of immunodiagnosis in certain circumstances.
