HBeAg在持续性HBV感染中的分子机制

StephenW

Hepatology International

January 2017, Volume 11, Issue 1, pp 79–86
Molecular mechanisms of HBeAg in persistent HBV infection

   
    Li-Min ChenXue-Gong Fan Email author Jing MaBo HeYong-Fang Jiang Email author

    Li-Min Chen        12
    Xue-Gong Fan        1 Email author
    Jing Ma        3
    Bo He        3
    Yong-Fang Jiang        3 Email author

    1.Infectious Diseases InstituteThe Xiangya Hospital, Central South UniversityChangshaPeople’s Republic of China
    2.Infectious Diseases InstituteThe Third Xiangya Hospital, Central South UniversityChangshaPeople’s Republic of China
    3.Liver Diseases Research CenterThe Second Xiangya Hospital, Central South UniversityChangshaPeople’s Republic of China

Original Article

First Online:
    18 May 2016

DOI: 10.1007/s12072-016-9734-5

Cite this article as:
    Chen, LM., Fan, XG., Ma, J. et al. Hepatol Int (2017) 11: 79. doi:10.1007/s12072-016-9734-5

    198 Downloads

Abstract
Purpose

The aim of this study is to investigate the T-lymphocyte subpopulation and expression of programmed cell death-1 (PD-1), toll-like receptor (TLR)3, TLR4, and interferon (INF)-γ to illustrate the relationship between hepatitis B e antigen (HBeAg) and persistent hepatitis B virus (HBV) infection.
Methods

Blood was taken from normal subjects into anticoagulation tubes to separate peripheral blood mononuclear cells (PBMCs). The PBMCs were divided into four groups and cultured with various concentrations of HBeAg for 72 h. Changes in the T-cell subset were analyzed through cell counting by flow cytometry, and expression of TLR3, TLR4, and PD-1 was assessed by flow cytometry and Western blot. The concentration of IFN-γ was analyzed using enzyme-linked immunospot (ELISPOT) experiments.
Results

PBMCs were stimulated with various concentrations of HBeAg for 72 h and assayed by flow cytometry to determine CD4+ and CD8+ cell counts. The relative frequencies of CD4+ and CD8+ subpopulations and the CD4+/CD8+ ratio decreased compared with the control group, and T-cell impairment was significantly associated with higher HBeAg load. TLR3, TLR4, and PD-1 protein expression was assessed using flow cytometry and Western blotting. Expression of TLR3, TLR4, and PD-1 increased with increasing concentration of HBeAg. ELISPOT experiments were used to determine the concentration of IFN-γ. IFN-γ production in treatment groups was lower than in the control group. Comparing IFN-γ production in treatment groups, IFN-γ production in PBMCs stimulated with high dose of HBeAg was lower than for those stimulated with low-dose HBeAg.
Conclusions

HBeAg can inhibit proliferation of lymphocytes, increase TLR3, TLR4, and PD-1 expression, and decrease IFN-γ production. This may be one of the molecular mechanisms of HBV immune tolerance.
Keywords
Hepatitis B e antigen Toll-like receptor 3 Toll-like receptor 4 Programmed cell death-1 Interferon-γ

X.-G. Fan and Y.-F. Jiang are equal contributors.

StephenW

肝病学国际

2017年1月，第11卷，第1期，第79-86页
HBeAg在持续性HBV感染中的分子机制

   
    李敏摘要：摘要：摘要：

    李敏民12
    Xue-Gong Fan 1电子邮件作者
    静马3
    Bo He 3
    Yong-Fang Jiang 3电子邮件作者

    1.中南大学湘雅医院感染性疾病研究所。长沙中华人民共和国
    2.中南大学第三湘雅医院感染性疾病研究所长沙中华人民共和国
    3.肝病研究中心中南大学第二湘雅医院中华人民共和国

原文

首先在线：
    2016年5月18日

DOI：10.1007 / s12072-016-9734-5

引用本文：
    Chen，LM。，Fan，XG。，Ma，J.et al。 Hepatol Int（2017）11：79.doi：10.1007 / s12072-016-9734-5

    198下载

摘要
目的

这项研究的目的是调查T淋巴细胞亚群和程序性细胞死亡-1（PD-1），toll​​样受体（TLR）3，TLR4和干扰素（INF）-γ的表达，以说明之间的关系乙型肝炎e抗原（HBeAg）和持续性乙型肝炎病毒（HBV）感染。
方法

将血液从正常受试者取入抗凝血管以分离外周血单核细胞（PBMC）。将PBMC分成四组，并用不同浓度的HBeAg培养72小时。通过流式细胞术通过细胞计数分析T细胞亚群中的变化，并通过流式细胞术和Western印迹评估TLR3，TLR4和PD-1的表达。使用酶联免疫斑点（ELISPOT）实验分析IFN-γ的浓度。
结果

用不同浓度的HBeAg刺激PBMC 72小时，并通过流式细胞术测定以测定CD4 +和CD8 +细胞计数。 CD4 +和CD8 +亚群的相对频率和CD4 + / CD8 +比与对照组相比降低，并且T细胞损伤与更高的HBeAg负荷显着相关。 TLR3，TLR4和PD-1蛋白表达用流式细胞术和Western印迹进行评估。 TLR3，TLR4和PD-1的表达随着HBeAg浓度的增加而增加。 ELISPOT实验用于测定IFN-γ的浓度。治疗组IFN-γ产生低于对照组。比较治疗组中IFN-γ产生，用高剂量HBeAg刺激的PBMC中的IFN-γ产生低于用低剂量HBeAg刺激的那些。
结论

HBeAg可以抑制淋巴细胞的增殖，增加TLR3，TLR4和PD-1的表达，降低IFN-γ的产生。这可能是HBV免疫耐受的分子机制之一。
关键词
乙型肝炎e抗原Toll样受体3 Toll样受体4程序性细胞死亡-1干扰素-γ

X.-G. Fan和Y.-F.江是平等的贡献者。