皮膚假性淋巴瘤免疫組織化學(xué)和基因重排研究
發(fā)布時間：2019-05-20 02:19
【摘要】： 目的：皮膚假性淋巴瘤(cutaneous pseudolymphomas, CPL)在組織學(xué)上類似于皮膚惡性淋巴瘤,但臨床表現(xiàn)上卻為良性的生物學(xué)行為,其不完全符合皮膚惡性淋巴瘤的診斷標準。免疫組織化學(xué)檢測雖可鑒別其類型,但無法鑒別其良惡性,只能作為一種輔助性的診斷方法。本試驗中我們對CPL采用免疫組化二步法檢測CD3、CD20和CD45RO的表達情況,以期對CPL進行分型。并采用聚合酶鏈式反應(yīng)(polymerase chain reaction, PCR)檢測TCR-γ口IgH的基因重排情況,期望為CPL的臨床診斷提供參考。 方法：天津市長征醫(yī)院皮膚科病理室2002年3月—2008年7月保存完整且臨床和病理確診的CPL患者的石蠟組織28例,其中男14例,女14例,年齡21-80歲,平均年齡(46.60±18.72)歲。10例扁平苔蘚(lichen planus, LP)石蠟組織作對照,其中男7例,女3例,年齡17～66歲,平均年齡(43.52±12.75)歲。兩組患者性別構(gòu)成和年齡差異無統(tǒng)計學(xué)意義,具有可比性。對28例CPL和10例LP的石蠟包埋組織采用免疫組化染色法,應(yīng)用免疫組化二步法檢測皮損中CD3、CD20和CD45RO的表達情況。并以從石蠟包埋處理的皮膚活檢組織中提取的DNA為模板,應(yīng)用PCR／瓊脂糖凝膠電泳方法,檢測TCR-γ和IgH基因重排情況,同時以T細胞淋巴瘤和B細胞淋巴瘤標本為作為陽性對照。 實驗結(jié)果采用SPSS13.0統(tǒng)計軟件進行統(tǒng)計分析,免疫組化結(jié)果應(yīng)用方差分析,兩兩比較采用dunnett-t檢驗,基因重排結(jié)果采用Fisher確切概率法檢驗,以P0.05為差異有統(tǒng)計學(xué)意義。 結(jié)果：28例CPL組織病理顯示真皮淺層內(nèi)大量淋巴細胞浸潤,免疫組化結(jié)果顯示28例皮膚假性淋巴瘤皮損中以CD3及CD45RO陽性表達為主的共13例,為T細胞假性淋巴瘤；以CD20陽性表達為主的共15例,為B細胞假性淋巴瘤。28例CPL中TCR-γ基因重排時,有8例出現(xiàn)smear帶,其余20例重排均為陰性,無1例重排陽性。對照組10例LP中有3例出現(xiàn)smear帶,其余7例重排均陰性,無1例重排陽性。20例CPL標本與10例LP石蠟包埋標本,其TCR-γ基因重排的多克隆率經(jīng)Fisher確切概率法檢驗,差異無統(tǒng)計學(xué)意義(P=0.615)；28例CPL中IgH基因重排時,有1例出現(xiàn)smear帶,其余27例重排均為陰性,無1例重排陽性。對照組10例LP中有1例出現(xiàn)smear帶,其余9例重排均陰性,無1例重排陽性。20例CPL標本與10例LP石蠟包埋標本,其IgH基因重排的多克隆率經(jīng)Fisher確切概率法檢驗,差異無統(tǒng)計學(xué)意義(P=0.462)。 結(jié)論：皮膚假性淋巴瘤組織真皮淺層內(nèi)大量淋巴細胞浸潤,主要是表達了CD3、CD45RO的T細胞,和表達CD20的B細胞；CPL組織內(nèi)淋巴細胞不存在明顯的TCR-γ基因和IgH基因重排,與生物學(xué)良性的扁平苔蘚皮損比較沒有明顯的差異；提示TCR-γ基因和IgH基因多克隆重排檢測在鑒別CPL良惡性方面具有參考意義。
[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 鹵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 鹵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- 緯 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- 緯 gene rearrangement was tested by Fisher exact probability method. There was no significant difference (P 鈮，
本文編號：2481267
[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 鹵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 鹵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- 緯 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- 緯 gene rearrangement was tested by Fisher exact probability method. There was no significant difference (P 鈮，
本文編號：2481267
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