Detection of clonal T-cell receptor gamma gene rearrangements in early mycosis fungoides/Sezary syndrome by polymerase chain reaction and denaturing gradient gel electrophoresis (PCR/DGGE).
Detection of clonal T-cell receptor gamma gene rearrangements in early mycosis fungoides/Sezary syndrome by polymerase chain reaction and denaturing gradient gel electrophoresis (PCR/DGGE).
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通过聚合酶链反应和变性梯度凝胶电泳 (PCR/DGGE) 检测早期蕈样肉芽肿/Sezary 综合征中克隆 T 细胞受体 γ 基因重排。
DOI：
10.1111/1523-1747.ep12389114
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发表时间：
1994
期刊：
影响因子：
--
通讯作者：
Heald,P
中科院分区：
文献类型：
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作者：
Wood,GS;Tung,RM;Haeffner,AC;Crooks,CF;Liao,S;Orozco,R;Veelken,H;Kadin,ME;Koh,H;Heald,P
我们使用基因扩增策略分析了185例T细胞受体(TCR)基因重排，包括真菌样肉芽肿/Sezary综合征(MF/SS)、其他皮肤肿瘤、炎症性皮肤病、反应性淋巴组织和正常皮肤。从皮损组织中提取基因组DNA，用V-γ1-8或V-γ9基因片段重排特异的引物，用聚合酶链式反应扩增TcR-γ链基因重排。用变性梯度凝胶电泳法(DGGE)将扩增产物按核苷酸序列和大小进行分离。68例MF/SS患者中有61例检测到显性克隆性TcR-γ基因重排。通过TCR基因重排的Southern印迹分析，在这些相同的病例中，有17例寻求显性克隆时，敏感性为90%，而当寻求显性克隆性时，敏感性仅为59%。这种敏感度的差异在早期浸润性皮肤病变中最大。在检测两例早期MF患者外周血侵犯方面，PCR/DGGE也比Southern印迹法更敏感。在另外12例疑似MF/SS患者中，9例(75%)经聚合酶链式反应/凝胶电泳法检测到TcR-γ基因克隆性重排，其中6例已确诊为MF/SS，3例为未确诊的MF/SS。在10 5例非MF/SS标本中，仅6例(6%)检测到显性TcR-γ基因重排。4例诊断为慢性皮炎，2例诊断为皮肤淋巴组织增生症。我们认为绝大多数MF/SS病例，包括斑片期疾病，都具有显性克隆性TcR-γ基因重排。在诊断为MF/SS的患者中，PCR/DGGE比Southern印迹分析更敏感地检测显性克隆和疾病分期。然而，由于PCR/DGGE对检测慢性皮炎患者亚群中显性TCR-y基因重排足够敏感，因此不能将其作为建立T细胞淋巴瘤诊断的唯一标准。与其他分子生物学克隆一样，在一些组织学上非特异性皮炎的病例中，克隆可以识别以前未识别的患者亚群，即那些患有克隆性皮炎的患者。在这些患者中确定MF/SS的长期风险是很重要的，因为我们的研究表明，MF/SS有时会出现与克隆性皮炎难以区分的皮损。
We used a gene amplification strategy to analyze T-cell receptor (TCR) gene rearrangements in 185 specimens, including mycosis fungoides/Sezary syndrome (MF/SS), other cutaneous neoplasms, inflammatory dermatoses, reactive lymphoid tissues,and normal skin. Genomic DNA was extracted from lesional tissues and rearrangements of the TCR-γ chain gene were amplified using the polymerase chain reaction (PCR) with primers specific for rearrangements involving Vγ1-8 or Vγ9 gene segments. The resulting PCR products were then separated according to their nucleotide sequence as well as size by denaturing gradient gel electrophoresis (DGGE). Dominant clonal TCR-γ gene rearrangements were detected in 61 of 68 MF/SS cases by PCR/DGGE. This sensitivity of 90% compared to a sensitivity of only 59% when dominant clonality was sought in 17 of these same cases by Southern blot analysis of TCR-& gene rearrangements. This difference in sensitivity was greatest in early, minimally infiltrated skin lesions. PCR/DGGE was also more sensitive than Southern blot analysis for detecting peripheral blood involvement in two cases of early MF. Among 12 additional specimens of suspected MF/SS, nine (75%) showed clonal TCR-γ gene rearrangements by PCR/DGGE including six of eight cases with a previously confirmed diagnosis of MF/SS and three of four cases without prior known MF/SS. Among 105 non-MF/SS specimens, dominant TCR-γ gene rearrangements were detected in only six cases (6%). Four were diagnosed as chronic dermatitis and two were diagnosed as cutaneous lymphoid hyperplasia.We conclude that the large majority of MF/SS cases, including patch phase disease, possess dominant clonal TCR-γ gene rearrangements. PCR/DGGE is more sensitive than Southern blot analysis for detecting dominant clonality and staging disease in patients with a confirmed diagnosis of MF/SS. However, because PCR/DGGE is sensitive enough to detect dominant TCR-y gene rearrangements in a subset of patients with chronic dermatitis, it cannot be used as the sole criterion for establishing a diagnosis of T-cell lymphoma. As with other molecular biologic clonality in some cases of histologically nonspecific dermatitis allows the identification of a previously unrecorgnized subset of patients, i.e., those with “clonal dermatitis”. It will be important to determine the long-term risk of MF/SS among these patients because our study indicated that MF/SS can sometimes present with lesions indistinguishable from clonal dermatitis.