Li Xuefen, Tian Li, Dong Yuejiao, Zhu Qiaoyun, Wang Yiyin, Han Wenzheng, Liu Xia, Ni Qin, Chen Yu, Li Lanjuan
State Key Laboratory for Diagnosis and Treatment of Infectious Diseases, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, The First Affiliated Hospital, College of Medicine, Zhejiang University, 79 Qingchun Road, Hangzhou 310003, PR China Department of Laboratory Medicine, The First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou 310003, PR China.
Department of Laboratory Medicine, The First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou 310003, PR China.
Clin Sci (Lond). 2015 Sep;129(5):395-404. doi: 10.1042/CS20140511. Epub 2015 Apr 14.
Interleukin (IL)-35 is an inhibitory cytokine consisting of IL-12A and Epstein-Barr virus-induced gene 3 (Ebi3) and is required by regulatory T-cells (Tregs) for maximal activity. During chronic hepatitis B virus (HBV) infection, Tregs have immunosuppressive effects on HBV-specific T helper (Th) cells, yet little is known about the complex regulation of Tregs and their contribution to the inadequate immune system response to the virus. In the present study, we investigated whether IL-35 is involved in HBV-related cellular immune responses. Cluster of differentiation (CD)4(+) T-cells from peripheral blood were derived from healthy volunteers, resolved HBV individuals and chronic active hepatitis B patients and stimulated with CD3/28-conjugated beads. We analysed mRNA and protein levels of IL-35 and assessed the inhibitory effect of IL-35 on HBV core antigen-specific cytotoxic T lymphocytes (CTLs), dendritic cells (DCs) and effector T-cells (Teffs). Correlation analyses between liver inflammation and HBV DNA load were conducted. Results show that chronic HBV patients harbour significantly higher levels of Ebi3 mRNA and protein in CD4(+) T-cells compared with healthy volunteers and resolved HBV individuals. IL-35 suppressed the proliferation of HBV antigen-specific CTLs and interferon (IFN)-γ production in vitro. Ex vivo, IL-35 decreased the proliferation of CD4(+)CD45RA(+) naïve T-cells, especially in CD4(+)CD25(-)CD45RA(+) naïve Teffs. IL-35 inhibited the expansion of CD11c(+) DCs. Our data indicate that IL-35 is highly expressed in chronic HBV CD4(+) T-cells and plays an important role in the inhibition of the cellular immune response in chronic HBV.
白细胞介素（IL）-35是一种由IL-12A和EB病毒诱导基因3（Ebi3）组成的抑制性细胞因子，调节性T细胞（Tregs）需要它来发挥最大活性。在慢性乙型肝炎病毒（HBV）感染期间，Tregs对HBV特异性辅助性T（Th）细胞具有免疫抑制作用，但关于Tregs的复杂调节及其对病毒免疫反应不足的贡献知之甚少。在本研究中，我们调查了IL-35是否参与HBV相关的细胞免疫反应。从健康志愿者、已康复的HBV感染者和慢性活动性乙型肝炎患者的外周血中分离出分化簇（CD）4(+) T细胞，并用CD3/28偶联磁珠进行刺激。我们分析了IL-35的mRNA和蛋白水平，并评估了IL-35对HBV核心抗原特异性细胞毒性T淋巴细胞（CTLs）、树突状细胞（DCs）和效应T细胞（Teffs）的抑制作用。进行了肝脏炎症与HBV DNA载量之间的相关性分析。结果显示，与健康志愿者和已康复的HBV感染者相比，慢性HBV患者的CD4(+) T细胞中Ebi3 mRNA和蛋白水平显著更高。IL-35在体外抑制了HBV抗原特异性CTLs的增殖和干扰素（IFN）-γ的产生。在体外，IL-35降低了CD4(+)CD45RA(+)初始T细胞的增殖，尤其是在CD4(+)CD25(-)CD45RA(+)初始Teffs中。IL-35抑制了CD11c(+) DCs的扩增。我们的数据表明，IL-35在慢性HBV CD4(+) T细胞中高表达，并在慢性HBV细胞免疫反应抑制中起重要作用。
