Clonality of cutaneous B-cell infiltrates determined by microdissection and immunoglobulin gene rearrangement
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S Signoretti1, M Murphy, P Puddu
1Department of Pathology, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts 02115, USA.
Insights
Detecting B-cell monoclonality in primary cutaneous B-cell lymphoma (PCBCL) is crucial for diagnosis. Microdissection enhances the sensitivity of immunoglobulin heavy chain (IgH) gene-rearrangement analysis by polymerase chain reaction (PCR), improving diagnostic accuracy.
Area of Science:
- Dermatopathology
- Molecular Diagnostics
- Oncology
Background:
- Accurate diagnosis of primary cutaneous B-cell lymphoma (PCBCL) relies on identifying a monoclonal B-cell population.
- Immunoglobulin heavy chain (IgH) gene-rearrangement analysis via polymerase chain reaction (PCR) is a key method for detecting B-cell monoclonality in tissue samples.
- Interpreting PCR results can be challenging due to the presence of reactive B lymphocytes in PCBCL specimens.
Purpose of the Study:
- To evaluate the effectiveness of IgH gene-rearrangement analysis using PCR on whole tissue sections versus microdissected B-cell subpopulations for diagnosing PCBCL.
- To determine if microdissection improves the sensitivity of PCR-based clonality assays in challenging PCBCL cases.
- To assess the utility of this technique in classifying unclassified lymphoid infiltrates (ULI).
Main Methods:
- IgH gene-rearrangement analysis by PCR was performed on DNA extracted from whole tissue sections of 19 cutaneous lymphoid infiltrates (10 PCBCL, 9 ULI).
- For specimens negative for monoclonality on whole sections, PCR analysis was repeated on microdissected B-cell subpopulations.
- Results were analyzed to compare the detection rates of monoclonal bands between whole tissue and microdissection approaches.
Main Results:
- Analysis of whole tissue sections detected monoclonality in 40% of PCBCL cases and showed non-specific patterns in all ULI cases.
- Subsequent microdissection identified monoclonal bands in an additional 30% of PCBCL cases, increasing the overall detection rate to 70%.
- Microdissection also identified a dominant clone in one ULI case, suggesting potential early PCBCL.
Conclusions:
- Microdissection significantly enhances the sensitivity of PCR-based IgH gene-rearrangement analysis for diagnosing PCBCL compared to whole tissue analysis.
- This refined technique aids in detecting B-cell monoclonality in challenging cases and may help identify early-stage PCBCL within ULI.
- Microdissection improves the diagnostic yield of clonality assays in cutaneous B-cell lymphomas.
Abstract:
Diagnosis of primary cutaneous B-cell lymphoma (PCBCL) is supported by the demonstration of a monoclonal B-cell population. Immunoglobulin heavy chain (IgH) gene-rearrangement analysis by polymerase chain reaction (PCR) is a reliable technique to detect B-cell monoclonality in paraffin-embedded tissue, but the presence of numerous reactive B lymphocytes in PCBCL may complicate the interpretation of clonality test results. To test this hypothesis, IgH gene-rearrangement analysis by PCR was performed on paraffin-embedded whole tissue sections of 19 cutaneous B-cell infiltrates diagnosed either as consistent with PCBCL (10 specimens) or unclassified lymphoid infiltrates (ULI) (9 specimens). In specimens that did not show monoclonal bands by IgH gene-rearrangement on DNA extracted from whole tissue sections, clonality assays were repeated on microdissected B-cell subpopulations suspicious for neoplastic cells. In the analysis of whole tissue sections, 4 (40%) of 10 specimens consistent with PCBCL showed one or two monoclonal bands, whereas 9 of 9 ULI specimens showed either a ladder or a smear. Clonality analysis of microdissected B-cell subpopulations showed 3 additional PCBCL specimens (total, 7 of 10) and 1 ULI specimen (total, 1 of 9) with unequivocal and reproducible monoclonal bands. Addition of microdissection increases the sensitivity of PCR-based B-cell clonality assay in PCBCL compared with analysis performed on the whole section (70% versus 40% monoclonal cases) and allows the recognition of a dominant clone in ULI specimens, possibly representing early PCBCL.
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