EASL2015:各种模式识别受体（PRR）直接抗病毒效果激动剂HBV复

StephenW

RS-1641 Viral hepatitis Hepatitis B & D - Experimental DIRECT ANTIVIRAL EFFECTS OF VARIOUS  PATTERN RECOGNITION RECEPTOR (PRR) AGONISTS IN HBV-REPLICATING HEPATOCYTES Julie Lucifora1, 2, Sarah Maadadi2, 3, Océane Floriot1, 2, Stephane Daffis4, Simon Fletcher4, Fabien Zoulim1, 2, 5, David Durantel* 1, 2 1U1052, INSERM, 2UCBL, University of Lyon, 3INSERM, U1052, Lyon, France, 4Gilead Sciences, Foster City, United States, 5Liver unit, Hospices Cilvils de Lyon, Lyon, France Corresponding author’s email: [email protected] Background and Aims: Current therapies for chronic hepatitis B virus (HBV) infection are effective at suppressing viral replication and improve long-term outcome, but have only low rates of HBsAg loss and anti-HBs seroconversion. There is an urgent need to identify new antiviral therapies to achieve functional cure and so decrease the risk of end-stage liver disease. Studies with the oral TLR7 agonist GS-9620 in animal models of chronic hepatitis B (CHB) have recently highlighted the potential of therapeutic immune modulation by small molecule agonists of innate immunity. To further explore the potential of PRR agonists to induce efficient and durable immune control against HBV, we tested the direct anti-HBV effect of different PRR agonists in cell culture models of natural infection. Methods: Primary human hepatocytes (PHH) and differentiated HepaRG cells (dHepaRG), which are both innate immune competent cells, were infected with HBV for 7 days, and then treated with various concentrations of different PRR agonists for 7 days. Antiviral activity was evaluated by quantifying HBeAg and HBsAg secretion (ELISA), HBV RNA (qRT-PCR) and total intracellular HBV DNA and cccDNA (qPCR), and toxicity measured by MTS assay and ApoB ELISA. Cytokine production was evaluated by ELISA 24 hours (h) post-stimulation. Results: The cytokines IP-10 and/or IL-6 were secreted by both PHH and dHepaRG 24h after stimulation with TLR1/2, TLR3, TLR4, TLR5, TLR2/6, RIG-I/MDA5, and AIM2 agonists, which correlated with a strong and dose-dependent antiviral effect in the absence of toxicity. The maximal antiviral effect was induced by Pam3CSK4 (i.e. TLR1/2-L), which induced sustained reduction in all viral parameters, including levels of cccDNA. Stimulation with TLR7, TLR8 and TLR9 agonists did not significantly induce IP-10 or IL-6 production and, with the exception of imiquimod, did not inhibit HBV replication. Interestingly, the antiviral response induced by imiquimod was independent of innate immune activation, suggesting an additional mode of action that should be further investigated. Conclusions: Our data highlight the potential of direct innate immunity activation in hepatocytes in the control of HBV replication, and support the recent interest in the development of PRR-based antiviral strategies against HBV. In addition, our data suggest that elucidation of the mode of antiviral action of imiquimod may lead to the identification of a novel target for the treatment of CHB.   Disclosure of Interest: J. Lucifora:  Grant: Gilead, S. Maadadi: : None Declared, O. Floriot: : None Declared, S. Daffis:  Employee: Gilead Sciences, S. Fletcher:  Employee: Gilead Sciences, F. Zoulim:  Grant: Gilead Sciences, D. Durantel:  Grant: Gilead Sciences

        

StephenW

RS-1641

病毒性肝炎

乙肝＆D - 实验





各种模式识别受体（PRR）直接抗病毒效果激动剂HBV复制的肝细胞

朱莉Lucifora1，2，莎拉Maadadi2，3，Océane酒店Floriot1，2，斯特凡Daffis4，西蒙Fletcher4，法比安斯基Zoulim1，2，5，大卫Durantel * 1，2

1U1052，INSERM，2UCBL，里昂，3INSERM，U1052，法国里昂，4Gilead科学，福斯特市，美国，5Liver单位，收容所Cilvils里昂，法国里昂大学



通讯作者的邮箱：[email protected]



背景和目的：目前治疗慢性乙型肝炎病毒（HBV）感染是在抑制病毒复制，改善长期预后效果，但是有HBsAg消失和抗-HBs阳转率只有很低。因此迫切需要确定新的抗病毒疗法来实现功能治愈等降低终末期肝病的风险。研究与口腔TLR7激动剂的GS-9620在慢性乙型肝炎（CHB）的动物模型，最近强调的治疗免疫调节的潜能由先​​天免疫的小分子激动剂。进一步探讨PRR激动剂的潜在诱导抗HBV高效耐用免疫控制，我们测试了不同的PRR激动剂的自然感染的细胞培养模型的直接抗HBV效果。

方法：原代人肝细胞（PHH）和分化HepaRG细胞（dHepaRG），这两者都是天然免疫能力的细胞，感染了HBV的7天，然后用不同浓度的不同的PRR激动剂的治疗7天。抗病毒活性通过定量HBeAg和HBsAg的分泌（ELISA），乙型肝炎病毒的RNA（QRT-PCR）和总的细胞内HBV DNA和cccDNA的（定量PCR）进行评估，并且通过MTS测定和载脂蛋白B ELISA测量毒性。细胞因子的产生，通过ELISA 24小时（八）刺激后评估。

结果：细胞因子IP-10和/或IL-6由两个PHH和dHepaRG 24小时的刺激与TLR1 / 2，TLR3，TLR4，TLR5，TLR2 / 6，RIG-I / MDA5后分泌，和AIM2激动剂，其中相关与在不存在毒性强而剂量依赖性抗病毒效果。最大的抗病毒效应诱导Pam3CSK4（即TLR1 / 2-L），从而诱发持续减少所有的病毒参数，包括cccDNA的水平。刺激TLR7，TLR8和TLR9的激动剂没有显著诱导的IP-10或IL-6的产生和，除咪喹莫特，不抑制HBV复制。有趣的是，诱导咪喹莫特的抗病毒反应是独立的先天免疫激活，提示作用的附加模式应作进一步调查。

结论：我们的数据直接突出先天免疫激活的肝细胞中的潜在于HBV复制的控制，以及支持在的抗HBV PRR-基于抗病毒策略的发展，最近的兴趣。此外，我们的数据表明，咪喹莫特的抗病毒作用的方式澄清可能导致一种新的目标识别为CHB的治疗。



股权变动：J. Lucifora：格兰特：Gilead公司，S. Maadadi：无申报，O. Floriot：无申报，S. Daffis：员工：Gilead Sciences公司，S.弗莱彻：员工：Gilead Sciences公司，F. Zoulim：格兰特：吉利德科学，D. Durantel：格兰特：Gilead Sciences公司

luckable

请问stephenw，抑制剂，激动剂分别表示什么意思，有什么不同。我们现在吃的这些抗病毒药物是什么剂？

StephenW

回复 luckable 的帖子

我解释只是一般性意见, 没有技术意识.

激动剂(agonist)是一种化学物质，与受体结合并激活该受体以产生生物学响应。而激动剂引起的动作，拮抗剂( antagonist)阻断激动剂的作用.

激动剂(agonist), 拮抗剂( antagonist)涉及受体(receptor)
抑制剂涉及酶(enzyme)
抑制剂(inhibitor)不相关拮抗剂(antagonist) 或激动剂(agonist).

抗病毒药物是抑制剂.

luckable

StephenW 发表于 2015-4-18 17:40
回复 luckable 的帖子

我解释只是一般性意见, 没有技术意识.

解释的很清楚。那就是激动剂很有未来，毕竟抑制不是终极目标。

newchinabok

gs9620     2015年9月（最终数据收集日期为主要测量指标）

zgct

，

咬牙硬挺

解释的很好啊，赞