Characterization of Feline Immunoglobulin Heavy Chain Variable Region Genes for the Molecular Diagnosis of B-cell Neoplasia
Open Access
- 1 September 2005
- journal article
- Published by SAGE Publications in Veterinary Pathology
- Vol. 42 (5), 596-607
- https://doi.org/10.1354/vp.42-5-596
Abstract
To develop a molecular-based assay so that the diagnosis of feline B-cell neoplasia can be facilitated, we have characterized 24 feline immunoglobulin heavy chain variable region (IGH V) complementary DNA (cDNA) transcripts. Structural homology with rearranged human IGH V genes was found, and the sequence information was used to design a feline-specific polymerase chain reaction (PCR)-based assay to amplify the complementarity determining region 3 as a marker for B-cell clonality. Conserved primers derived from the second and third framework regions of V gene segments were used in conjunction with 2 sequence-specific primers and 1 degenerate primer derived from the J gene segments. Each PCR reaction was run in duplicate, and both native and denatured PCR products were evaluated using polyacrylamide gel electrophoresis. Formalin-fixed, paraffin-embedded (FFPE) tissue sections from cats with confirmed B-cell neoplasia (diffuse large B-cell lymphoma, plasmacytoma, and myeloma) were examined, and 15/22 (68.2%) cats produced results indicative of the presence of a monoclonal population of B cells. The evaluation of denatured PCR products (heteroduplex analysis) facilitated a more accurate interpretation in 3/15 (20%) cats. Pseudoclonality was a major reason for the failure to detect monoclonality. Poor DNA quality is a significant concern and was responsible for the removal of 2 cats from the study. Using this assay, FFPE normal feline lymphoid tissues and unfixed peripheral blood mononuclear cells were determined to be composed of polyclonal populations of B cells. This assay represents a useful adjunctive diagnostic tool for the diagnosis and investigation of feline B-cell lymphoproliferative disorders.Keywords
This publication has 17 references indexed in Scilit:
- Design and standardization of PCR primers and protocols for detection of clonal immunoglobulin and T-cell receptor gene recombinations in suspect lymphoproliferations: Report of the BIOMED-2 Concerted Action BMH4-CT98-3936Leukemia, 2003
- Histopathologic and Immunophenotypic Characterization of Extramedullary Plasmacytomas in Nine CatsVeterinary Pathology, 2003
- Diagnosis of Canine Lymphoid Neoplasia Using Clonal Rearrangements of Antigen Receptor GenesVeterinary Pathology, 2003
- Fusion gene transcripts and Ig/TCR gene rearrangements are complementary but infrequent targets for PCR-based detection of minimal residual disease in acute myeloid leukemiaLeukemia, 2002
- Sensitive Detection of Clonal Immunoglobulin Rearrangements in Frozen and Paraffin Embedded Tissues by Polymerase Chain Reaction Heteroduplex AnalysisDiagnostic Molecular Pathology, 2000
- Limitations of clonality analysis of B cell proliferations using CDR3 polymerase chain reactionMolecular Pathology, 2000
- Specificity of PCR-based clonality analysis of immunoglobulin heavy chain gene rearrangements for the detection of bone marrow involvement by low-grade B-cell lymphomasThe Journal of Pathology, 2000
- Molecular characterization of illegitimate TCRδ gene rearrangements in acute myeloid leukaemiaBritish Journal of Haematology, 1994
- Improved PCR method for detecting monoclonal immunoglobulin heavy chain rearrangement in B cell neoplasms.Journal of Clinical Pathology, 1992
- Development of a highly sensitive assay, based on the polymerase chain reaction, for rare B‐lymphocyte clones in a polyclonal populationBritish Journal of Haematology, 1990