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Updated: Aug 9, 2026
Isolating And Immunostaining Lymphocytes and Dendritic Cells from Murine Peyer's Patches
Published on: March 17, 2013
Reliable and cost-effective paraffin section immunohistology of lymphoproliferative disorders
G H Segal1, M H Stoler, A J Fishleder
1Department of Pathology, Cleveland Clinic Foundation, Ohio.
Insights
A panel of three monoclonal antibodies (CD20, CD43, CD45RO) efficiently screens lymphoid proliferations in paraffin sections. This immunotyping method accurately assigns lineage for most B-cell and T-cell lymphomas.
Area of Science:
- Hematopathology
- Immunohistochemistry
- Molecular Pathology
Background:
- Accurate lineage assignment of lymphoid proliferations is crucial for diagnosis and treatment.
- Conventional immunotyping methods may require fresh tissue, which is not always available.
- Paraffin-embedded tissues offer a valuable resource for retrospective diagnostic studies.
Purpose of the Study:
- To evaluate a three-antibody panel (CD20, CD43, CD45RO) for immunotyping lymphoid proliferations in paraffin-embedded tissues.
- To assess the utility of this panel as an efficient and cost-effective screening tool.
- To compare immunohistochemistry results with conventional immunotyping and genotyping.
Main Methods:
- Studied 110 neoplastic and reactive lymphoid proliferations using B5-fixed, paraffin-embedded tissues.
- Employed three monoclonal antibodies: CD20 (L26), CD43 (Leu22), and CD45RO (UCHL1).
- Compared immunohistochemistry results with conventional immunotyping and Southern blot hybridization for genotyping in 54 cases.
Main Results:
- Lineage assignment by paraffin immunohistology matched conventional methods in 73/76 B-cell and all 5 T-cell tumors.
- CD20 reacted with 73/76 B-cell tumors; CD43 showed coexpression with CD20 in 11/12 cases.
- CD43 and CD45RO marked T-cell lymphomas effectively.
- Paraffin immunohistology and genotyping concordance was observed in 48/50 cases.
Conclusions:
- The CD20, CD43, and CD45RO antibody panel is an efficient and cost-effective method for immunotyping lymphoid proliferations in paraffin sections.
- This panel aids in accurate lineage assignment for most B-cell and T-cell lymphomas.
- Fresh or frozen tissue may still be necessary for resolving ambiguous cases and profiling phenotypic deletions.
Abstract:
We studied 110 neoplastic and reactive lymphoid proliferations with three monoclonal antibodies--CD20 (L26), CD43 (Leu22), and CD45RO (UCHL1)--on B5-fixed, paraffin-embedded tissue to evaluate the utility of this panel as an immunotypic screen of such lesions. All cases were initially immunotyped by conventional methods. Genotyping by Southern blot hybridization was also done in 54 cases. Seventy-four of 79 malignant lymphomas and both of two hairy cell leukemias were of B-cell origin; and five lymphomas were defined as T-cell lineage. Lineage assignment was identical for paraffin section immunohistology and conventional immunotyping in 73 of 76 B cell and all of five T-cell tumors. CD20 was reactive with 73 of 76 B-cell tumors. CD43 was reactive with 12 of 74 B-cell lymphomas, and CD20/CD43 coexpression was seen in 11 of these cases. CD43 and CD45RO marked all of five and three of five T-cell lymphomas, respectively. Lineage assignment was identical for paraffin immunohistology and genotyping in 48 of 50 cases with identifiable gene rearrangements. Twenty-four nonneoplastic and five Hodgkin's disease cases that were studied also showed similar immunoreactivity patterns by both paraffin and conventional immunotypic methods. This panel of three monoclonal antibodies is an efficient, cost-effective approach for immunotyping most lymphoid proliferations in paraffin sections. Nevertheless, the pathologist should always try to obtain fresh or frozen tissue to aid in resolving occasional discrepant cases, to establish clonality in morphologically ambiguous ones, and to profile prognostically important phenotypic deletions.
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