基础医学与临床 ›› 2023, Vol. 43 ›› Issue (4): 621-625.doi: 10.16352/j.issn.1001-6325.2023.04.0621

• 研究论文 • 上一篇    下一篇

EBV裂解期蛋白BZLF1和BILF1抑制Raji细胞主要组织相容性复合体的表达

郑洪玲, 郭庆伟, 张伟, 张艳, 何兴波, 叶冰*   

  1. 山东大学附属儿童医院(济南市儿童医院)急诊科,山东 济南 250022
  • 收稿日期:2022-08-21 修回日期:2022-12-04 出版日期:2023-04-05 发布日期:2023-04-03
  • 通讯作者: *13583184890@163.com
  • 基金资助:
    济南市临床医学科技创新计划(202019071); 山东省医药卫生科技发展计划(202106010191)

EBV lytic proteins BZLF1 and BILF1 inhibit the expression of major histocompatibility complex in Raji cells

ZHENG Hongling, GUO Qingwei, ZHANG Wei, ZHANG Yan, HE Xingbo, YE Bing*   

  1. Department of Emergency, Children's Hospital Affiliated to Shandong University (Ji'nan Children's Hospital), Ji'nan 250022, China
  • Received:2022-08-21 Revised:2022-12-04 Online:2023-04-05 Published:2023-04-03
  • Contact: *13583184890@163.com

摘要: 目的 探讨EB病毒(EBV)裂解期蛋白BZLF1和BILF1是否能够抑制人B淋巴瘤细胞系Raji细胞主要组织相容性复合体(MHC)的表达。方法 用佛波酯(TPA)和丁酸钠(NaB)诱导Raji细胞促使EBV 激活进入裂解期,用Western bolt检测EBV裂解期的标志物BZLF1蛋白的表达; RT-qPCR检测EBV进入裂解期后BZLF1和BILF1基因的表达;流式细胞测量术检测Raji细胞MHC Ⅱ类分子和MHC Ⅰ类分子的表达。电穿孔转染质粒过表达BZLF1和BILF1蛋白及siRNA技术敲低BZLF1和BILF1蛋白表达后,分别用流式细胞术及Western bolt检测Raji细胞MHC Ⅱ类分子和MHC Ⅰ类分子的表达。结果 TPA/NaB 能够诱导EBV激活进入裂解期,EBV进入裂解期后BZLF1和BILF1基因过表达。EBV进入裂解期以及过表达BZLF1和BILF1后,MHC Ⅱ类分子和MHC Ⅰ类分子表达均明显降低(P<0.05); 特异性siRNA可以阻断BZLF1和BILF1对Raji细胞MHC Ⅱ类分子和MHC Ⅰ类分子的抑制作用(P<0.05)。 结论 EBV裂解期蛋白BZLF1和BILF1能够抑制Raji细胞MHC分子的表达。

关键词: BZLF1, BILF1, 主要组织相容性复合体 Ⅱ, 主要组织相容性复合体Ⅰ

Abstract: Objective To investigate whether BZLF1 and BILF1 can inhibit the expression of major histocompatibility complex(MHC)molecules in Raji cells during Epstein-Barr virus (EBV) lytic cycle. Methods Reactivation was triggered by treating Raji cells with TPA and NaB. As the marker protein during EBV lytic state, the BZLF1 protein expression was detected by Western blot. The expression of BZLF1 and BILF1 at transcription level was analyzed by RT-qPCR in EBV reactivation. The expression of MHC class Ⅱ and MHC class Ⅰ molecules was analyzed by flow cytometry. After BZLF1 and BILF1 over-expression or gene silencing via siRNA in Raji cells by electroporation, the expression of MHC class Ⅱ and MHC class Ⅰ molecules was detected by flow cytometry and Western blot respectively. Results EBV reactivation can be triggered with TPA and NaB. The BZLF1 and BILF1 gene expression was observed by inducing EBV reactivation in Raji cells. The levels of MHC class Ⅱ and MHC class Ⅰ molecules were down regulated by inducing EBV reactivation or ectopic expression of BZLF1 and BILF1 (P<0.05). Expression of MHC class Ⅱ and MHC class Ⅰ molecules was restored by specific siRNA. Conclusions BZLF1 and BILF1 down regulate the expression of MHC class Ⅱ and MHC class Ⅰ molecules in EBV reactivation respectively.

Key words: BZLF1, BILF1, major histocompatibility complex Ⅱ, major histocompatibility complex Ⅰ

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