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VDJ-Seq: Deep Sequencing Analysis of Rearranged Immunoglobulin Heavy Chain Gene to Reveal Clonal Evolution Patterns of B Cell Lymphoma
Published on: December 28, 2015
Molecular immunoglobulin/T- cell receptor clonality analysis in cutaneous lymphoproliferations. Experience with the
Yorick Sandberg1, Freerk Heule, King Lam
1Department of Immunology, Erasmus MC, University Medical Center Rotterdam, Rotterdam, The Netherlands.
Insights
BIOMED-2 multiplex PCR analysis reliably detects clonality in cutaneous B- and T-cell lymphomas (CBCL/CTCL). This molecular technique aids in diagnosing skin lymphomas and assessing extracutaneous spread when combined with clinical data.
Area of Science:
- Oncology
- Molecular Biology
- Dermatology
Background:
- Cutaneous lymphomas require accurate diagnosis to differentiate between benign and malignant conditions.
- Immunoglobulin (Ig) and T-cell receptor (TCR) gene clonality analysis is crucial for distinguishing lymphoproliferative skin lesions.
- Standardized molecular techniques are needed for reliable assessment of primary cutaneous lymphomas (CBCL/CTCL).
Purpose of the Study:
- To evaluate the BIOMED-2 multiplex polymerase chain reaction (PCR) strategy for Ig/TCR gene clonality analysis in suspected cutaneous lymphomas.
- To compare PCR-based methods with Southern blot (SB) for detecting clonality in CBCL and CTCL.
- To assess the utility of molecular clonality analysis in evaluating extracutaneous dissemination.
Main Methods:
- Studied 60 patients with suspected primary cutaneous B- or T-cell lymphoma (CBCL/CTCL).
- Assessed Ig/TCR gene rearrangement clonality using BIOMED-2 standardized PCR, Southern blot (SB), heteroduplex (HD), and GeneScan (GS) analysis.
- Compared molecular findings with histopathologic diagnoses based on EORTC classification.
Main Results:
- PCR detected Ig clonality in 83% of CBCL cases, while SB detected it in 92%.
- SB detected TCR clonality in 68% of CTCL cases; TCRG and TCRB PCR detected it in 76% and 66%, respectively.
- PCR GS analysis of TCR rearrangements was slightly more informative than HD analysis; clonality assessment aided in evaluating extracutaneous sites.
Conclusions:
- The BIOMED-2 multiplex PCR analysis is a reliable and useful diagnostic technique for suspected CBCL/CTCL.
- This molecular strategy aids in assessing extracutaneous dissemination of cutaneous lymphomas.
- Results should be interpreted alongside clinical, histologic, and immunophenotypic data for optimal diagnostic accuracy.
Background And Objectives:
Molecular clonality analysis of immunoglobulin (Ig) and T-cell receptor (TCR) genes is a widely used diagnostic tool for discrimination between polyclonal, oligoclonal, and monoclonal lymphoproliferative skin lesions. We studied Ig/TCR clonality in a series of 60 patients with an initial suspicion of (primary) cutaneous B- or T-cell lymphoma (CBCL/CTCL). Clonality of Ig/TCR gene rearrangements was assessed by Southern blot (SB) and polymerase chain reaction (PCR) analysis using standardized PCR primers and protocols of the BIOMED-2 Concerted Action BMH4-CT98-3936. The obtained PCR products were subjected to heteroduplex (HD) and GeneScan (GS) analysis. We compared the data of 154 samples with the histopathologic diagnosis, based on the EORTC classification of skin lymphomas.
Design And Methods:
Molecular results were largely concordant with histopathology. In 12 CBCL patients PCR analysis of Ig gene rearrangements detected clonality in 83% of cases whereas SB did so in 92%. Clonal TCR gene rearrangements were detected by SB in 68% of CTCL patients, whereas TCRG and TCRB PCR analysis detected clonality in 76% and 66% of cases, respectively. PCR GS analysis of TCR rearrangements appeared to be slightly more informative than HD analysis. Clonality assessment was particularly informative for studying involvement of extracutaneous sites, such as regional lymph nodes, peripheral blood, and bone marrow.
Interpretation And Conclusions:
Our study shows that the BIOMED-2 multiplex PCR analysis strategy is a reliable and useful technique in the diagnostic process of patients with an initial suspicion of (primary) CBCL/CTCL and for assessment of extracutaneous dissemination, provided that the results are interpreted in the context of clinical, histologic and immunophenotypic data.
