GCET2兔单克隆抗体的制备及其在弥漫性大B细胞淋巴瘤中初步应用  

Application of rabbit monoclonal antibody GCET2 in diagnosis of diffuse large B cell iymphoma

作　　者：潘红阳[1,2] 张昕霞 翁媛媛[1] 周韧[1] Pan Hongyang Zhang Xinxia Weng Yuanyuan Zhou Ren(Department of Pathology and Pathophysiology, Institute of Pathology and Forensic Medicine, Zhejiang University School of Medicine, Hangzhou 310058, China)

机构地区：[1]浙江大学病理学与法医研究所浙江大学病理学与病理生理学系,杭州310058 [2]宜康(杭州)生物技术有限公司

出　　处：《中华病理学杂志》2016年第12期844-849,共6页Chinese Journal of Pathology

摘　　要：目的开发制备GCET2兔单克隆抗体，研究其在弥漫性大B细胞淋巴瘤（DLBCL）诊断中的应用。方法采用兔单克隆抗体技术制备GCET2兔单克隆抗体；经免疫印迹、免疫组织化学（IHC）、免疫细胞化学和流式细胞术等方法鉴定其特异性；并用IHC法，检测GCET2与其他DLBCL诊断相关标志物（CD10、bcl-6、MUM1、GCET1、FOXP1、Ki-67和CMYC）在81例DLBCL、5例滤泡细胞淋巴瘤和2例Burkitt淋巴瘤临床样本组织中的表达。结果GCET2（EP316）抗体能特异性识别生发中心B细胞及其来源的肿瘤组织和细胞系（Raji、Daudi和Ramos）中的GCET2蛋白；对81例DLBCL进行检测，GCET2的阳性率达到43.2%（35/81），明显高于其他生发中心标志物的阳性检出率。此外，CMYC高表达DLBCL样本在GCET2阴性表达和阳性表达样本中所占比例分别为15.2%（7/46）和2.8%（1/35）。结论本研究制备的GCET2（EP316）抗体具有良好的特异性，是一种敏感的生发中心B细胞标记抗体。GCET2与其他分子标志物联用，可能有助于提高对生发中心B细胞型DLBCL的检测敏感性。Objective To prepare a rabbit monoelonal antibody GCET2 and to investigate its diagnostic value in the workup of diffuse large B-cell lymphoma （ DLBCL ） . Methods GCET2 rabbit monoclonal antibody was developed by using RabMAb technology, and its specificity was confirmed by ELISA, Western blot, immunohistoehemistry （IHC） and flow eytometry. A panel of immunomarkers including GCET2, CD10, bel-6, MUM1, GCET1, FOXP1 , Ki-67 and CMYC was evaluated in 81 cases of DLBCLs, 5 cases of follicular lymphomas （FL） and 2 cases of Burkitt＇s lymphomas. Results Rabbit monoelonal GCET2 antibody （clone EP316 ） was developed with specificity for normal germinal center B-cells （GCB） and GCB origined lymphomas. In 81 cases of DLBCL, the positive rate of GCET2 was 43.2% （35/81）, which was significantly higher than that of other germinal center markers. Moreover, among 81 DLBCLs, the proportions of high CMYC expression in GCET2 negative and positive groups were 15.2% （7/46） and 2. 8% （ 1/35 ）, respectively. Conclusion GCET2 is a sensitive immunohistochemical marker for GCB derived lymphomas. Combined with other biomarkers, it may improve the diagnostic sensitivity of GCB-DLBCL.

关 键 词：淋巴瘤 大B-细胞 弥漫性 生发中心 抗体 单克隆 

分 类 号：R733.1[医药卫生—肿瘤]