Andersen W K, Li N, Bhawan J
Department of Dermatology, Boston University School of Medicine, Massachusetts, USA.
J Cutan Pathol. 1999 Apr;26(4):176-82. doi: 10.1111/j.1600-0560.1999.tb01825.x.
Polymerase chain reaction (PCR)-based amplification of T-cell receptor (TCR)-gamma genes is a novel technique that can detect a clone of T cells comprising less than 1% of the total T cells in a lymphoid infiltrate. Besides greater sensitivity than Southern blotting, this technique can be performed with smaller quantities of lower molecular weight genomic DNA as template. We retrospectively analyzed 12 paraffin-embedded biopsies of cutaneous T-cell lymphoma (CTCL), 1 case suspicious for CTCL, 1 case of granulomatous slack skin, and 8 cases of inflammatory skin diseases to determine if PCR-denaturing gradient gel electrophoresis (PCR-DGGE) analysis can detect TCR-gamma gene rearrangements on paraffin-embedded specimens. We were able to amplify Vgamma1-8 TCR sequences in each case and detected a dominant clone in 9 of 12 cases of CTCL and in granulomatous slack skin. We analyzed Vgamma9 sequences in 9 cases of CTCL and detected a dominant clone in 4 cases. This study demonstrates that PCR-DGGE can easily be applied retrospectively to cutaneous biopsies of lymphoproliferative diseases when fresh tissue is not available.
基于聚合酶链反应(PCR)的T细胞受体(TCR)-γ基因扩增是一项新技术,它能够检测出在淋巴浸润中占总T细胞不到1%的T细胞克隆。除了比Southern印迹法具有更高的灵敏度外,该技术还可以使用更少量的低分子量基因组DNA作为模板进行操作。我们回顾性分析了12例皮肤T细胞淋巴瘤(CTCL)的石蜡包埋活检标本、1例疑似CTCL的病例、1例肉芽肿性皮肤松弛症病例以及8例炎症性皮肤病病例,以确定PCR-变性梯度凝胶电泳(PCR-DGGE)分析能否检测石蜡包埋标本上的TCR-γ基因重排。我们在每个病例中都成功扩增出了Vγ1-8 TCR序列,并在12例CTCL病例中的9例以及肉芽肿性皮肤松弛症病例中检测到了优势克隆。我们对9例CTCL病例分析了Vγ9序列,在4例中检测到了优势克隆。这项研究表明,当没有新鲜组织时,PCR-DGGE可以很容易地回顾性应用于淋巴增殖性疾病的皮肤活检。