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(American Journal of Pathology. 1999;155:355-363.)
© 1999 American Society for Investigative Pathology

Technical Advance

Detection of Immunoglobulin {kappa} Light Chain Rearrangements by Polymerase Chain Reaction

An Improved Method for Detecting Clonal B-CellLymphoproliferative Disorders

Jerry Z. Gong*{dagger}, Sherman Zheng*, Roberto Chiarle*, Christine De Wolf-Peeters{ddagger}, Giorgio Palestro{ddagger}§, Glauco Frizzera* and Giorgio Inghirami*{dagger}

From the Division of Hematopathology/Molecular Pathology Laboratory,*
Department of Pathology, and Kaplan Comprehensive Cancer Center,{dagger}
New York University School of Medicine, New York, New York; the Department of Pathology,{ddagger}
Leuven University, Leuven, Belgium; and the Department of Anatomic Pathology,{ddagger}
§
University of Torino, Torino, Italy

The clonal determination of B-cell lymphoproliferative disorders by immunoglobulin heavy chain (IgH) rearrangement by polymerase chain reaction (PCR) is widely used. However, few attempts have been made to detect immunoglobulin {kappa} light chain (Ig{kappa}) gene rearrangement using PCR. We studied 145 cases of B-cell neoplasms, along with 58 atypical and 18 reactive lymphoproliferative disorders, using newly designed degenerate oligoprimers recognizing the framework 3 (FR3{kappa}) and the joint (J{kappa}) regions of the Ig{kappa} gene. PCR products were analyzed on nondenaturing polyacrylamide gel (ndPAGE). Clonal B-cell determination was further investigated using IgH rearrangement and t(11:14) or t(14:18). By combining these methods, we detected either clonality or translocation in 117 of 137 cases (85%) in mature B-cell neoplasms. The additional analysis of Ig{kappa} rearrangement improved sensitivity from 66% to 85%. To investigate whether the Ig gene configuration could be characterized using Ig{kappa} PCR in B-cell neoplasms showing severe breakdown of genomic DNA, 18 selected cases were analyzed. Successful amplification was detected in 72% of the cases using either FR3/2-JH and/or FR3J{kappa} oligoprimers. Finally, clonality was detected in 21 of 58 atypical B-cell proliferations, and among them, the atypical marginal cell (54%) and atypical large cell (50%) proliferations showed the highest frequency of clonal immunoglobulin gene products. We concluded that PCR/ndPAGE analysis of Ig{kappa} is a sensitive, rapid, and efficient method for assessing clonality in conjunction with IgH and specific translocation analysis. This approach is particularly useful in the characterization of B-cell lymphoproliferative disorders in archival material with poor preservation of the genomic DNA.




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