Hayashi Y, Fukayama M, Funata N, Hishima T, Oba T, Koike M
Department of Pathology, Tokyo Metropolitan Komagome Hospital, Japan.
Pathol Int. 1999 Feb;49(2):110-7. doi: 10.1046/j.1440-1827.1999.00831.x.
Characterization of the clonality of non-Hodgkin's lymphoma (NHL) by the rearranged segments of immunoglobulin heavy chain (Ig(H)) or T cell receptor (TCR) genes is not only useful in the confirmation of the diagnosis but also for the future assessment of how a secondary lymphoma, such as a recurrence or another primary lymphoma, occurs. As a practical approach to obtaining and registering this information in a surgical pathology laboratory, FR3 and FR1 regions of Ig(H) gene and TCRgamma gene were concurrently amplified by polymerase chain reaction (PCR) using each pair of consensus primers and the same PCR protocol. Examined samples consisted of 134 primary NHL (phenotypically, 108 B cell and 26 T cell NHL), 19 reactive lymphadenopathies, as well as five secondary lymphomas whose primary lesions were included in this study. Among the primary NHL, the combined PCR analysis disclosed the clonality in 103 of 134 NHL (77%), by FR3 PCR in 77 B cell and two T cell NHL, by FR1 PCR in 59 B cell and one T cell NHL, and by TCRgamma PCR in 11 B cell and 17 of 26 T cell NHL, but in none of the reactive lymphadenopathies. Among the secondary lymphomas, the same pattern of PCR analysis was obtained in two cases (the durations between first and second lymphomas; 6 and 10 months), which suggested recurrence. In contrast, different results were obtained in three cases (17-37 months), which indicated another primary or emergence of the subclones. The results of Southern blot analysis were concordant with the PCR results of the first and the secondary lymphomas. Although the combined PCR analysis cannot replace Southern blot hybridization because of its lower detection rate, it can select those cases suitable for further Southern blot analysis thus reducing the number of unnecessary examinations by nearly 75%. This approach may also be useful in the comparative evaluation of primary and secondary lymphomas.
通过免疫球蛋白重链（Ig(H)）或T细胞受体（TCR）基因的重排片段来鉴定非霍奇金淋巴瘤（NHL）的克隆性，不仅有助于确诊，还可用于未来评估继发性淋巴瘤（如复发或另一种原发性淋巴瘤）的发生情况。作为在外科病理实验室获取和记录该信息的实用方法，使用每对共识引物并采用相同的PCR方案，通过聚合酶链反应（PCR）同时扩增Ig(H)基因和TCRγ基因的FR3和FR1区域。检测样本包括134例原发性NHL（表型上，108例B细胞和26例T细胞NHL）、19例反应性淋巴结病以及5例继发性淋巴瘤，其原发性病变包含在本研究中。在原发性NHL中，联合PCR分析显示134例NHL中有103例（77%）具有克隆性，通过FR3 PCR检测出77例B细胞和2例T细胞NHL，通过FR1 PCR检测出59例B细胞和1例T细胞NHL，通过TCRγ PCR检测出11例B细胞和26例T细胞NHL中的17例，而反应性淋巴结病均未检测出克隆性。在继发性淋巴瘤中，2例（首次和第二次淋巴瘤之间的间隔时间分别为6个月和10个月）获得了相同的PCR分析模式，提示复发。相比之下，3例（间隔时间为17 - 37个月）获得了不同结果，表明是另一种原发性淋巴瘤或亚克隆的出现。Southern印迹分析结果与原发性和继发性淋巴瘤的PCR结果一致。虽然联合PCR分析因其较低的检测率不能替代Southern印迹杂交，但它可以筛选出适合进一步进行Southern印迹分析的病例，从而将不必要检查的数量减少近75%。这种方法也可能有助于原发性和继发性淋巴瘤的比较评估。
