Kuroda H, Komatsu H, Nakamura S, Niitsu Y, Takahashi T, Ueda R, Seto M
Laboratory of Chemotherapy, Aichi Cancer Center Research Institute, Nagoya.
Jpn J Cancer Res. 1995 Sep;86(9):890-8. doi: 10.1111/j.1349-7006.1995.tb03102.x.
Recently, we produced a monoclonal antibody, 5D4, against the PRAD1/cyclin D1 product and suggested positive nuclear staining to be associated with mantle cell lymphoma (MCL). Now we have further characterized the specificity of this antibody and studied the relation of immunohistochemical detection to PRAD1/cyclin D1 mRNA expression and DNA rearrangement. Immunofluorescence and immunoblotting studies demonstrated the 5D4 antibody to be crossreactive with cyclin D2, but not cyclin D3. On immunostaining, 15 of 19 MCL cases (79%) presented the nuclear staining pattern and PRAD1/cyclin D1 mRNA expression was detected by Northern blot analysis in 12 of 15 MCL cases studied (80%): all cases with the mRNA expression showed the nuclear staining pattern. Southern blot analysis with 11q13 BCL-1 probes detected DNA rearrangements in 8 of 19 MCL cases (42%), all 8 exhibiting PRAD1/cyclin D1 mRNA expression. In 21 lymphoma cases of types other than MCL, neither the mRNA expression nor the nuclear staining were observed, although cytoplasmic staining was often apparent. These results indicated that positive nuclear staining of lymphoma cells by 5D4 antibody reflects PRAD1/cyclin D1 mRNA expression, and showed that this monoclonal antibody has diagnostic value for differentiating MCL from other types of lymphomas.
最近,我们制备了一种针对PRAD1/细胞周期蛋白D1产物的单克隆抗体5D4,并指出细胞核阳性染色与套细胞淋巴瘤(MCL)相关。现在我们进一步鉴定了该抗体的特异性,并研究了免疫组化检测与PRAD1/细胞周期蛋白D1 mRNA表达及DNA重排之间的关系。免疫荧光和免疫印迹研究表明,5D4抗体与细胞周期蛋白D2有交叉反应,但与细胞周期蛋白D3无交叉反应。在免疫染色中,19例MCL病例中有15例(79%)呈现细胞核染色模式,在15例研究的MCL病例中有12例(80%)通过Northern印迹分析检测到PRAD1/细胞周期蛋白D1 mRNA表达:所有有mRNA表达的病例均呈现细胞核染色模式。用11q13 BCL-1探针进行Southern印迹分析,在19例MCL病例中有8例(42%)检测到DNA重排,所有8例均有PRAD1/细胞周期蛋白D1 mRNA表达。在21例非MCL类型的淋巴瘤病例中,未观察到mRNA表达和细胞核染色,尽管细胞质染色常常很明显。这些结果表明,5D4抗体对淋巴瘤细胞的细胞核阳性染色反映了PRAD1/细胞周期蛋白D1 mRNA表达,并表明该单克隆抗体对鉴别MCL与其他类型淋巴瘤具有诊断价值。