皮肤假性淋巴瘤免疫组织化学和基因重排研究

发布时间：2019-05-20 02:19

【摘要】： 目的：皮肤假性淋巴瘤(cutaneous pseudolymphomas, CPL)在组织学上类似于皮肤恶性淋巴瘤,但临床表现上却为良性的生物学行为,其不完全符合皮肤恶性淋巴瘤的诊断标准。免疫组织化学检测虽可鉴别其类型,但无法鉴别其良恶性,只能作为一种辅助性的诊断方法。本试验中我们对CPL采用免疫组化二步法检测CD3、CD20和CD45RO的表达情况,以期对CPL进行分型。并采用聚合酶链式反应(polymerase chain reaction, PCR)检测TCR-γ口IgH的基因重排情况,期望为CPL的临床诊断提供参考。 方法：天津市长征医院皮肤科病理室2002年3月—2008年7月保存完整且临床和病理确诊的CPL患者的石蜡组织28例,其中男14例,女14例,年龄21-80岁,平均年龄(46.60±18.72)岁。10例扁平苔藓(lichen planus, LP)石蜡组织作对照,其中男7例,女3例,年龄17～66岁,平均年龄(43.52±12.75)岁。两组患者性别构成和年龄差异无统计学意义,具有可比性。对28例CPL和10例LP的石蜡包埋组织采用免疫组化染色法,应用免疫组化二步法检测皮损中CD3、CD20和CD45RO的表达情况。并以从石蜡包埋处理的皮肤活检组织中提取的DNA为模板,应用PCR／琼脂糖凝胶电泳方法,检测TCR-γ和IgH基因重排情况,同时以T细胞淋巴瘤和B细胞淋巴瘤标本为作为阳性对照。 实验结果采用SPSS13.0统计软件进行统计分析,免疫组化结果应用方差分析,两两比较采用dunnett-t检验,基因重排结果采用Fisher确切概率法检验,以P0.05为差异有统计学意义。 结果：28例CPL组织病理显示真皮浅层内大量淋巴细胞浸润,免疫组化结果显示28例皮肤假性淋巴瘤皮损中以CD3及CD45RO阳性表达为主的共13例,为T细胞假性淋巴瘤；以CD20阳性表达为主的共15例,为B细胞假性淋巴瘤。28例CPL中TCR-γ基因重排时,有8例出现smear带,其余20例重排均为阴性,无1例重排阳性。对照组10例LP中有3例出现smear带,其余7例重排均阴性,无1例重排阳性。20例CPL标本与10例LP石蜡包埋标本,其TCR-γ基因重排的多克隆率经Fisher确切概率法检验,差异无统计学意义(P=0.615)；28例CPL中IgH基因重排时,有1例出现smear带,其余27例重排均为阴性,无1例重排阳性。对照组10例LP中有1例出现smear带,其余9例重排均阴性,无1例重排阳性。20例CPL标本与10例LP石蜡包埋标本,其IgH基因重排的多克隆率经Fisher确切概率法检验,差异无统计学意义(P=0.462)。 结论：皮肤假性淋巴瘤组织真皮浅层内大量淋巴细胞浸润,主要是表达了CD3、CD45RO的T细胞,和表达CD20的B细胞；CPL组织内淋巴细胞不存在明显的TCR-γ基因和IgH基因重排,与生物学良性的扁平苔藓皮损比较没有明显的差异；提示TCR-γ基因和IgH基因多克隆重排检测在鉴别CPL良恶性方面具有参考意义。
[Abstract]:Objective: skin pseudolymphoma (cutaneous pseudolymphomas, CPL) is similar to skin malignant lymphoma in histology, but it is a benign biological behavior in clinical manifestation, which does not fully meet the diagnostic criteria of skin malignant lymphoma. Although Immunohistochemical detection can distinguish its type, it can not distinguish its benign and malignant, and can only be used as an auxiliary diagnostic method. In this study, we detected the expression of CD3,CD20 and CD45RO in CPL by two-step immunohistochemical method in order to classify CPL. The gene rearrangement of TCR- gamma mouth igh was detected by polymerase chain reaction (polymerase chain reaction, PCR), which was expected to provide a reference for the clinical diagnosis of CPL. Methods: from March 2002 to July 2008, 28 patients with CPL were preserved in the Department of Dermatology, Tianjin Changzheng Hospital, including 14 males and 14 females, aged 21 years and 80 years. The average age was (46.60 卤18.72) years. 10 cases of (lichen planus, LP) paraffin tissue of lichen planus were used as control, including 7 males and 3 females, 17 years old and 66 years old, with an average age of (43.52 卤12.75) years. There was no significant difference in gender composition and age between the two groups. The expression of CD3,CD20 and CD45RO in 28 cases of CPL and 10 cases of LP were detected by immunohistochemical staining. The rearrangement of TCR- 纬 and IgH genes was detected by PCR/ agarose gel electrophoresis using DNA extracted from paraffin embedded skin biopsies as template. At the same time, T-cell lymphomas and B-cell lymphomas were used as positive controls. The experimental results were statistically analyzed by SPSS13.0 statistical software, the results of immunohistochemistry were analyzed by variance analysis, the pairwise results were compared by dunnett-t test, and the results of gene rearrangement were tested by Fisher exact probability method, and the difference was statistically significant (P 0.05). Results: in 28 cases of CPL, a large number of lymphocytes infiltrated in the superficial dermis. The results of immunohistochemistry showed that the positive expression of CD3 and CD45RO was mainly in the lesions of 28 cases of skin pseudolymphoma, which was T cell pseudolymphoma. A total of 15 cases were mainly CD20 positive, which was B cell pseudolymphoma. In 28 cases of CPL, 8 cases had smear band, the other 20 cases were negative, and no rearrangement was positive in 28 cases of CPL. In the control group, 3 cases of LP showed smear band, the other 7 cases were negative and no rearrangement was positive. 20 cases of CPL specimens and 10 cases of LP paraffin embedded specimens, the polyclonal rate of TCR- 纬 gene rearrangement was tested by Fisher exact probability method. There was no significant difference (P 鈮，

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