Tümkaya T, Comans-Bitter W M, Verhoeven M A, van Dongen J J
Department of Immunology, Erasmus University Rotterdam, University Hospital Dijkzigt, The Netherlands.
Leukemia. 1995 Dec;9(12):2127-32.
Southern blot analysis of immunoglobulin (Ig) genes has proven to be important for detection of clonal rearrangements in patients with lymphoproliferative diseases. To improve the detection of clonal Ig lambda (Ig lambda) gene rearrangements, we carefully determined the precise restriction map of the J-C lambda gene region, developed a suitable C lambda probe (IGLC3), and evaluated relevant restriction enzymes in combination with the IGLC3 probe. For the latter purpose, we selected 75 B cell malignancies with proven expression of Ig lambda protein chains in order to be sure that each malignancy contained at least one clonally rearranged IG lambda allele. Our extensive Southern blot analyses with the IGLC3 probe in EcoRI and/or HindIII digests revealed that combined EcoRI/HindIII digestion detected Ig lambda gene rearrangements in 95% of the 75 patients and 94% of the 98 rearranged alleles. In contrast, HindIII and EcoRI single digests allowed detection of rearrangements in only 78% and 83% of the patients, and 67% and 79% of rearranged alleles, respectively. We conclude that the use of the IGLC3 probe in combined EcoRI/HindIII digests is superior to EcoRI and HindIII single digests. This probe/enzyme combination is informative for clonality studies in approximately 95% of patients.
免疫球蛋白（Ig）基因的Southern印迹分析已被证明对于检测淋巴增殖性疾病患者的克隆重排很重要。为了提高克隆性Igλ（Igλ）基因重排的检测率，我们仔细确定了J-Cλ基因区域的精确限制性图谱，开发了合适的Cλ探针（IGLC3），并评估了与IGLC3探针联合使用的相关限制性酶。为了实现后一个目的，我们选择了75例已证实表达Igλ蛋白链的B细胞恶性肿瘤，以确保每个恶性肿瘤至少包含一个克隆性重排的IGλ等位基因。我们用IGLC3探针在EcoRI和/或HindIII消化物中进行的广泛Southern印迹分析表明，EcoRI/HindIII联合消化在75例患者中的95%和98个重排等位基因中的94%检测到了Igλ基因重排。相比之下，HindIII和EcoRI单消化分别仅在78%和83%的患者以及67%和79%的重排等位基因中检测到重排。我们得出结论，在EcoRI/HindIII联合消化中使用IGLC3探针优于EcoRI和HindIII单消化。这种探针/酶组合对大约95%的患者的克隆性研究具有参考价值。
