Gong J Z, Zheng S, Chiarle R, De Wolf-Peeters C, Palestro G, Frizzera G, Inghirami G
Division of Hematopathology/Molecular Pathology Laboratory, Department of Pathology, Kaplan Comprehensive Cancer Center, New York, USA.
Am J Pathol. 1999 Aug;155(2):355-63. doi: 10.1016/s0002-9440(10)65132-2.
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 (Igkappa) 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 (FR3kappa) and the joint (Jkappa) regions of the Igkappa 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 Igkappa rearrangement improved sensitivity from 66% to 85%. To investigate whether the Ig gene configuration could be characterized using Igkappa 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 FR3Jkappa 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 Igkappa 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.
通过聚合酶链反应（PCR）检测免疫球蛋白重链（IgH）重排来进行B细胞淋巴增殖性疾病的克隆性判定已被广泛应用。然而，利用PCR检测免疫球蛋白κ轻链（Igκ）基因重排的尝试却很少。我们使用新设计的简并寡核苷酸引物，识别Igκ基因的框架3（FR3κ）和连接（Jκ）区域，研究了145例B细胞肿瘤，以及58例非典型和18例反应性淋巴增殖性疾病。PCR产物在非变性聚丙烯酰胺凝胶（ndPAGE）上进行分析。使用IgH重排和t(11;14)或t(14;18)进一步研究克隆性B细胞判定。通过结合这些方法，我们在137例成熟B细胞肿瘤中的117例（85%）中检测到了克隆性或易位。Igκ重排的额外分析将敏感性从66%提高到了85%。为了研究在显示基因组DNA严重降解的B细胞肿瘤中，是否可以使用Igκ PCR来表征Ig基因构型，我们分析了18例选定的病例。使用FR3/2-JH和/或FR3Jκ寡核苷酸引物，在72%的病例中检测到了成功的扩增。最后，在58例非典型B细胞增殖中检测到21例克隆性，其中非典型边缘细胞增殖（54%）和非典型大细胞增殖（50%）显示出克隆性免疫球蛋白基因产物的最高频率。我们得出结论，Igκ的PCR/ndPAGE分析是一种与IgH和特异性易位分析相结合来评估克隆性的敏感、快速且有效的方法。这种方法在表征基因组DNA保存不佳的存档材料中的B细胞淋巴增殖性疾病时特别有用。
