AASLD2016[600]干扰素-α是早期病毒抑制的关键介质 在乙型肝炎

StephenW

600
Interferon-Alpha is a key mediator of early viral suppression
in HBeAg negative variants of Hepatitis B virus
Zina Valaydon1,2, Alexander Thompson1, Paul V. Desmond1, Peter
A. Revill3, Stephen Locarnini3, Marc Pellegrini2, Gregor Ebert2;
1Gastroenterology, St Vincent’s Hospital, Melbourne, VIC, Australia;
2Walter and Eliza Hall Institute, Melbourne, VIC, Australia;
3VIDRL, Melbourne, VIC, Australia
Background: The precore antigen,HBeAg,is thought to be a
tolerogen in Hepatitis B virus (HBV) infection.However the
underlying mechanisms are unknown.The basal core promoter
(BCP) and precore (PC) variants of HBV reduce and
abolish HBeAg production respectively and are associated
with increased risk of cirrhosis and liver cancer.We used a
novel immunocompetent mouse model of HBV infection to study
differences between wild type (WT) virus and BCP/PC HBV
variants that express little or no HBeAg. Aims: To characterize
the viral kinetics and immunopathogenesis of BCP/PC
variants using a mouse model of HBV. Methods: An HBV 1.2
overlength genome was flanked by inverted terminal repeats in
a plasmid vector and mutated using site-directed mutagenesis
to introduce dual A1762T/G1764A BCP and G1896A PC
mutations.WT, PC and BCP plasmids were hydrodynamically
injected in the tail vein of C57/BL6 mice.The role of interferon-
alpha (IFNa), interferon-gamma (IFNg) and tumour necrosis
factor-alpha (TNFa) were investigated using IFNa receptor
knockout (IFNaR-KO) mice, IFNg-KO mice and TNFa neutralizing
antibodies respectively. Results:Significant differences in
viral kinetics were seen with the BCP and PC mutants vs. WT
(Fig1a).There was an initial rapid drop in viral load (VL)in the
mutants in the early phase of infection(p<0.03 at week 2-7).
To interrogate the phenomenon a series of immune experiments
were performed.When the experiment was repeated in
IFNaR-KO mice,there was a significant increase in VL in PC
and BCP mutants (p<0.05) but no effect in the WT.The early
difference in VL between mutant and WT strains was abolished
when the influence of IFNa was removed (Fig 1b).In contrast,
there was no difference in IFNg-KO mice and mice treated with
TNFa neutralizing antibodies, suggesting that neither TNFa nor
IFNg is responsible for the early viral suppression in mutants.
Conclusions: Our data suggests that IFNa is a key mediator of
the early innate immune response and we propose that HBeAg
is an IFN resistance protein that enables chronicity. Complementation
studies are being carried out to confirm this.
Fig 1a: Differences in VL in BCP/PV vs WT (n=15-17) and
Fig 1b: Loss of VL differences when experiment repeated in
IFNaR-KO mice (n=8)
Disclosures:
Alexander Thompson - Advisory Committees or Review Panels: Gilead Sciences,
Abbvie, BMS, Merck / MSD; Grant/Research Support: Gilead Sciences, Abbvie,
BMS, Merck / MSD; Speaking and Teaching: Gilead Sciences, Abbvie, BMS,
Merck / MSD, Roche Diagnostics
Peter A. Revill - Grant/Research Support: Gilead Sciences
Stephen Locarnini - Consulting: Gilead Sciences Inc, Arrowhead Research Corp,
Spring Bank Pharmaceuticals, Inc.; Employment: Melbourne Health
Marc Pellegrini - Consulting: TetraLogic Pharmaceuticals
The following people have nothing to disclose: Zina Valaydon, Paul V. Desmond,
Gregor Ebert

StephenW

AASLD2016 [600]干扰素-α是早期病毒抑制的关键介质
在乙型肝炎病毒的HBeAg阴性变体中
干扰素-α是早期病毒抑制的关键介质
在乙型肝炎病毒的HBeAg阴性变体中
Zina Valaydon1,2，Alexander Thompson1，Paul V. Desmond1，Peter
A. Revill3，Stephen Locarnini3，Marc Pellegrini2，Gregor Ebert2;
1胃肠病学，圣文森特医院，墨尔本，澳大利亚维多利亚州;
2Walter和Eliza Hall Institute，Melbourne，VIC，Australia;
3VIDRL，Melbourne，VIC，Australia
背景：前列腺抗原，HBeAg，被认为是
耐受原在乙型肝炎病毒（HBV）感染
基础核心启动子
（BCP）和前核（PC）变体
废除HBeAg生产分别和相关联
增加肝硬化和肝癌的风险
新型免疫小鼠模型的HBV感染研究
野生型（WT）病毒和BCP / PC HBV之间的差异
表达很少或不表达HBeAg的变体。目的：表征
BCP / PC的病毒动力学和免疫发病机制
使用HBV的小鼠模型的变体。方法：
超长基因组的侧翼是反向末端重复序列
质粒载体并使用定点诱变进行突变
介绍双A1762T / G1764A BCP和G1896A PC
WT，PC和BCP质粒进行流体动力学分析
注射在C57 / BL6小鼠的尾静脉。干扰素 -
α（IFNa），干扰素-γ（IFNg）和肿瘤坏死
因子-α（TNFa）
敲除（IFNaR-KO）小鼠，IFNg-KO小鼠和TNFα中和
抗体。结果：
使用BCP和PC突变体相对于WT观察到病毒动力学
（图1a）。病毒载量（VL）的初始快速下降
突变体在感染的早期（在第2-7周p <0.03）。
要询问现象一系列免疫实验
当实验重复进行时
IFNαR-KO小鼠中，PC中的VL有显着增加
和BCP突变体（p <0.05），但在WT中无效
突变株和WT株之间的VL差异被消除
当去除IFNα的影响时（图1b）。相比之下，
IFNg-KO小鼠和用其处理的小鼠没有差异
TNFa中和抗体，表明既不是TNFa也不是
IFNg负责突变体中的早期病毒抑制。
结论：我们的数据表明IFNa是一个关键调解人
早期先天免疫反应，我们建议HBeAg
是允许长期性的IFN抗性蛋白。互补
正在进行研究以确认这一点。
图1a：BCP / PV与WT（n = 15-17）和VL中VL的差异
图1b：重复实验时VL差异的损失
IFNaR-KO小鼠（n = 8）
披露：
Alexander Thompson - 咨询委员会或审查小组：G​​ilead Sciences，
Abbvie，BMS，Merck / MSD;资助/研究支持：Gilead Sciences，Abbvie，
BMS，Merck / MSD;讲座和教学：吉利德科学，Abbvie，BMS，
Merck / MSD，Roche Diagnostics
Peter A. Revill - 资助/研究支持：吉利德科学
Stephen Locarnini - 咨询：吉利德科学公司，Arrowhead研究公司，
春天药业有限公司就业：墨尔本健康
Marc Pellegrini - 咨询：TetraLogic制药
以下人士没有透露：Zina Valaydon，Paul V. Desmond，
Gregor Ebert