Objective: To investigate the TNFAIP3/A20 abnormalities and its association with Epstein-Barr virus (EBV) in classical Hodgkin lymphoma (CHL). Methods: Formalin-fixed, paraffinembedded tissue blocks of 54 CHL patients were collected and subjected to the construction of tissue microarray (TMA) for further analyses. EBV status was evaluated by in situ hybridization (ISH) for EBER1/2 and immunohistochemistry (IHC) with anti-LMP-1 antibody. Fluorescence in situ hybridization (FISH) and IHC were performed to determine the copy number alterations of TNFAIP3 and A20 protein expression respectively. Results: The concordance rate of IHC for LMP-1 and ISH for EBER1/2 was100%, and 25.9% (14/54) cases were identified with EBV infection. Immunohistochemistry analysis demonstrated 27.8% (15/54) cases with A20 expression deficiency. Of the 54 cases tested for A20 expression, 49 cases were simultaneously analyzed by FISH, which showed 10 (20.4% ) cases harboring TNFAIP3 deletion. However, discrepancy was observed between the results of A20 by IHC and TNFAIP3 deletion by FISH. Only 1 case with TNFAIP3 deletion demonstrated complete loss of A20 immunoreactivity. In addition, comparison of the frequency of either A20 expression loss or TNFAIP3 deletion between EBV-positive and-negative cases did not reveal any significance (P>0.05). Conclusion: TNFAIP3 deletion could be observed in both EBV-positive and - negative CHL cases. A20 expression by IHC could not confirm TNFAIP3 deletion by FISH, which might be related to the technical issues.
目的: 研究经典霍奇金淋巴瘤（CHL）中肿瘤坏死因子α诱导蛋白3(TNFAIP3）基因及其编码的A20蛋白表达情况，并分析其与Epstein-Barr病毒（EBV）感染的相关性。
方法: 收集54例CHL患者的病理标本及临床资料，选取肿瘤细胞丰富区制作组织芯片。应用免疫组织化学染色法检测EBV编码的潜伏膜蛋白1(LMP-1），原位杂交法检测EBV编码的RNA（EBER1/2）以明确EBV感染状态。使用位点特异性间期荧光原位杂交（FISH）法检测TNFAIP3基因表达，免疫组织化学染色法检测A20蛋白表达。使用SPSS17.0统计学软件包进行数据分析。
结果: LMP-1、EBER1/2阳性率均为25.9%（14/54），两者符合率为100%。27.8%（15/54）的标本存在A20表达丢失，20.4%（10/49）的标本存在TNFAIP3杂合或纯合缺失。两种方法的检测结果显示存在明显不一致，TNFAIP3缺失伴A20丢失仅有1例。此外，EBV感染阴性与A20丢失及TNFAIP3缺失无相关性（P>0.05）。
结论: EBV阳性和阴性CHL病例均存在TNFAIP3基因和（或）A20蛋白表达丢失。FISH和免疫组织化学染色两种方法结果不一致，可能与技术因素有关。