Ben-Ezra J
Department of Pathology, City of Hope National Medical Center, Duarte, CA 91010.
Leuk Lymphoma. 1992 Jul;7(4):289-95. doi: 10.3109/10428199209049780.
The use of clonospecific probes has recently been employed for the detection of minimal residual disease in B- and T-cell acute lymphoblastic leukemias. However, these methods are predicated upon the successful amplification of the V-D-J rearrangement in the genome of the tumor cells by the polymerase chain reaction (PCR). In order to determine whether the type of B-cell lymphoid malignancy influenced the rate of success of amplifying the region of the immunoglobulin heavy chain gene rearrangement in these lesions, we studied 41 morphologically and immunologically well characterized B-cell neoplasms. DNA was extracted from frozen tissue of the lymphomas and leukemias, and subjected to PCR amplification using a 5' immunoglobulin heavy chain gene variable region consensus Framework 3 region (FR3) primer, and a 3' consensus primer for the immunoglobulin heavy chain joining region. One or two distinct bands, representing the rearranged immunoglobulin heavy chain gene, were detected in six of six small non-cleaved cell lymphomas, five of five small lymphocytic lymphomas, four of six acute lymphoblastic leukemias, four of six follicular lymphomas, three of six diffuse mixed small and large cell lymphomas, one of six diffuse large cell lymphomas, and one of six immunoblastic large cell lymphomas; all control cases of lymphocyte predominant Hodgkin's disease (5/5) and reactive follicular hyperplasia (5/5) were negative. We therefore conclude that the type of B-cell neoplasm influences the ability to detect immunoglobulin gene rearrangements by PCR with currently used consensus primers.
克隆特异性探针最近已被用于检测B细胞和T细胞急性淋巴细胞白血病中的微小残留病。然而,这些方法基于通过聚合酶链反应(PCR)成功扩增肿瘤细胞基因组中的V-D-J重排。为了确定B细胞淋巴瘤的类型是否会影响在这些病变中扩增免疫球蛋白重链基因重排区域的成功率,我们研究了41例形态学和免疫学特征明确的B细胞肿瘤。从淋巴瘤和白血病的冷冻组织中提取DNA,并使用5'免疫球蛋白重链基因可变区共有框架3区(FR3)引物和免疫球蛋白重链连接区的3'共有引物进行PCR扩增。在6例小无裂细胞淋巴瘤中的6例、5例小淋巴细胞淋巴瘤中的5例、6例急性淋巴细胞白血病中的4例、6例滤泡性淋巴瘤中的4例、6例弥漫性混合小细胞和大细胞淋巴瘤中的3例、6例弥漫性大细胞淋巴瘤中的1例以及6例免疫母细胞性大细胞淋巴瘤中的1例中检测到一条或两条代表重排免疫球蛋白重链基因的不同条带;所有淋巴细胞为主型霍奇金病(5/5)和反应性滤泡增生(5/5)的对照病例均为阴性。因此,我们得出结论,B细胞肿瘤的类型会影响使用当前使用的共有引物通过PCR检测免疫球蛋白基因重排的能力。