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Updated: Aug 26, 2025
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
A clonality assay in canine B cell tumors targeting the immunoglobulin light chain lambda locus
Masamine Takanosu1, Yumiko Kagawa2
1Nasunogahara Animal Clinic, Ohtawara, Tochigi, Japan.
Insights
Canine clonality assays can now utilize the immunoglobulin light chain lambda (IGL) gene. This new method accurately detects lymphoma in dogs, showing high sensitivity for diffuse large B cell lymphoma.
Area of Science:
- Veterinary immunology
- Molecular diagnostics
- Canine oncology
Background:
- Clonality assays for antigen receptor rearrangement aid in diagnosing lymphoproliferative diseases.
- Previous canine clonality assays focused on immunoglobulin heavy chain (IGH) and immunoglobulin light chain kappa (IGK) loci.
- The immunoglobulin light chain lambda (IGL) locus has not been previously investigated for canine clonality.
Purpose of the Study:
- To evaluate the diagnostic utility of clonality assays targeting the IGL locus in dogs.
- To assess the sensitivity and specificity of IGL-based clonality assays in canine lymphomas and non-lymphoma samples.
Main Methods:
- Genomic DNA was extracted from formalin-fixed paraffin-embedded tissues of canine diffuse large B cell lymphomas (DLBCL), cutaneous plasmacytomas, and non-lymphoma lymph nodes.
- Sequences of IGLV and IGLJ gene segments were obtained, and primers were designed for polymerase chain reaction (PCR).
- PCR products were analyzed using capillary electrophoresis to determine clonality.
Main Results:
- IGL-based clonality assays detected clonality in 20 out of 23 DLBCL cases (87.0%).
- Clonality was observed in 8 out of 30 cutaneous plasmacytomas (26.7%).
- Specificity was high at 95.7%, with only one non-lymphoma lymph node showing clonality.
Conclusions:
- The IGL locus is a viable target for canine clonality assays.
- IGL-based assays demonstrate high sensitivity for DLBCL but lower sensitivity for cutaneous plasmacytomas.
- These findings expand the molecular tools available for diagnosing canine lymphoproliferative diseases.
Abstract:
Clonality assays for antigen receptor rearrangement have been used as adjunct examinations of lymphoproliferative diseases. These assays have been useful for differentiation between inflammation and clonal expansion of lymphocytes. Whereas the immunoglobulin heavy chain (IGH) and immunoglobulin light chain kappa (IGK) loci have been targeted in canine clonality assays previously, the immunoglobulin light chain lambda gene (IGL) locus has not yet been investigated. This study aimed to evaluate the usefulness of clonality assays in dogs using IGL. Canine diffuse large B cell lymphomas (DLBCL), cutaneous plasmacytomas, and pathologically diagnosed lymph nodes without lymphoma, were used in this study. Genomic DNA was extracted from formalin-fixed paraffin embedded sections. Sequences of IGLV and IGLJ were obtained from the ImMunoGeneTics database. Several primers against IGLVs and IGLJs were designed in the regions showing homology, by alignment of the gene segments. Products of polymerase chain reaction were analyzed on a capillary electrophoresis. In total, 20 of 23 cases of DLBCL showed clonality (87.0 %), whereas 8 of 30 cutaneous plasmacytomas were clonal (26.7 %). One of 23 lymph nodes without lymphoma showed clonality, thus the specificity was 95.7 %. These data indicate that the IGL locus could be a target for canine clonality assays and that the sensitivity of IGL-based clonality assays in cutaneous plasmacytomas was lower than that in DLBCL.
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