Martinelli G, Testoni N, Farabegoli P, Zaccaria A, Amabile M, Visani G, Pelliconi S, Raspadori D, Manfroi S, Vittone A, Terragna C, Tura S
Institute of Hematology L. e A. Seràgnoli, Ospedale S. Orsola University of Bologna, Italy.
Leuk Lymphoma. 1997 Jun;26(1-2):131-9. doi: 10.3109/10428199709109168.
We applied a simple polymerase chain reaction (PCR) based method for detecting immunoglobulin heavy-chain (IgH) gene rearrangement, using its CDR-III region to assess B-cell clonality in a series of 100 acute lymphoblastic leukemias (ALL) (84 B-cell lineage, 4 null-ALL and 12 T-ALL). The amplified CDR-III regions can be generated in all the B-lineage ALL and separated by size by polyacrylamide gel electrophoresis (PAGE), thereby providing a specific diagnostic marker for each B cell clone. Size heterogeneity resulting from independent IgH rearrangement events and the high resolution power after electrophoresis and silver staining of the PAGE gels can be used to generate a "fingerprint" of the PCR fragments representing either the spectrum of B-cell clonality in complex populations of B lymphocytes or the partially genomic configuration of the VH-N-DH-N-JH region. At diagnosis, we found the presence of one clonal IgH heavy-chain gene rearrangement in 80 B-cell lineage and null ALL and a biclonal rearrangement in 8 cases. The CDR-III bands were of sizes ranging from 80 to 130 bp. The PCR analysis of the IgH gene enabled us to obtain a CDR-III leukemia specific product in all cases, thereby providing a specific and diagnostic marker for each B-cell clone.
我们应用了一种基于简单聚合酶链反应(PCR)的方法来检测免疫球蛋白重链(IgH)基因重排,利用其互补决定区III(CDR-III)区域评估100例急性淋巴细胞白血病(ALL)(84例B细胞系、4例无丙种球蛋白血症ALL和12例T细胞ALL)中的B细胞克隆性。在所有B细胞系ALL中均可产生扩增的CDR-III区域,并通过聚丙烯酰胺凝胶电泳(PAGE)按大小分离,从而为每个B细胞克隆提供特异性诊断标志物。由独立的IgH重排事件导致的大小异质性以及PAGE凝胶电泳和银染后的高分辨率,可用于生成代表B淋巴细胞复杂群体中B细胞克隆性谱或VH-N-DH-N-JH区域部分基因组构型的PCR片段“指纹”。在诊断时,我们发现80例B细胞系和无丙种球蛋白血症ALL中存在一种克隆性IgH重链基因重排,8例存在双克隆重排。CDR-III条带大小在80至130 bp之间。对IgH基因的PCR分析使我们能够在所有病例中获得CDR-III白血病特异性产物,从而为每个B细胞克隆提供特异性诊断标志物。