Abou-Elella A, Shafer M T, Wan X Y, Velanker M, Weisenburger D D, Nathwani B N, Gascoyne R D, Greiner T C, Chan W C
Department of Pathology and Microbiology, University of Nebraska Medical Center, Omaha 68198-3135, USA.
Am J Clin Pathol. 2000 Oct;114(4):516-22. doi: 10.1309/X559-FDJB-LJGD-YG7E.
We investigated the clonal relationship between follicular center cell and monocytoid B-cell components of non-Hodgkin lymphoma by isolating the components and comparing the nucleotide sequences of the complementarity-determining region (CDR)3 of the rearranged immunoglobulin heavy chain (IgH) gene. Paraffin blocks from 4 cases with amplifiable DNA using the polymerase chain reaction (PCR) were identified. Multiple representative cell clusters of the 2 components were obtained by microdissection, and the IgH CDR3 was amplified using a seminested PCR. Most of the PCR products obtained from both tumor components in each case had identical lengths when analyzed with polyacrylamide gel electrophoresis (PAGE) and identical migratory patterns on denaturing gradient gel electrophoresis (DGGE). These findings indicate sequence identity of the IgH CDR3 of both tumor components. Sequence analysis showed that point mutations were responsible for bands from the same case that had nonidentical migratory patterns by DGGE. The components in each of the 4 cases studied have the same clonal origin. Intraclonal sequence variations in the IgH gene were observed in 2 cases, consistent with the presence of continued somatic hypermutation after establishment of the clone. The expression of CD10 and bcl-2, as well as the detection of bcl-2 rearrangements in 2 cases, indicate that these lymphomas are of follicular center cell origin.
我们通过分离非霍奇金淋巴瘤的滤泡中心细胞和单核样B细胞成分，并比较重排的免疫球蛋白重链（IgH）基因互补决定区（CDR）3的核苷酸序列，研究了它们之间的克隆关系。利用聚合酶链反应（PCR）从4例可扩增DNA的石蜡块中进行鉴定。通过显微切割获得这两种成分的多个代表性细胞簇，并用半巢式PCR扩增IgH CDR3。用聚丙烯酰胺凝胶电泳（PAGE）分析时，每个病例中从两种肿瘤成分获得的大多数PCR产物长度相同，在变性梯度凝胶电泳（DGGE）上迁移模式相同。这些结果表明两种肿瘤成分的IgH CDR3序列相同。序列分析表明，点突变导致了同一病例中DGGE迁移模式不同的条带。所研究的4例中的每例成分都有相同的克隆起源。在2例中观察到IgH基因的克隆内序列变异，这与克隆形成后持续的体细胞超突变的存在一致。2例中CD10和bcl-2的表达以及bcl-2重排的检测表明，这些淋巴瘤起源于滤泡中心细胞。
