EASL2012:PegINFN-alppha alone or in combination with entecavir in mice

StephenW

Abstract               
                                

Title	Pegylated-interferon-alpha alone or in combination with entecavir restores ISG responsiveness and reduces intrahepatic viral loads and antigenemia in hepatitis B virus infected humanized mice
Speaker:	Lena   Allweiss
Author:	L. Allweiss1*, M. Lütgehetmann1,2, T. Volz1, T. Bornscheuer1, A.W. Lohse1, J. Petersen3, H. Ma4, K. Klumpp4, S. Fletcher4, M. Dandri1
Affiliation:	1Internal Medicine, 2Medical Microbiology, Virology & Hygiene, University Medical Center Hamburg-Eppendorf, 3Liver Centre, Asklepiosklinik St. Georg, Hamburg, Germany, 4Hoffmann-La Roche, Nutley, NJ, USA. *[email protected]
Background: The direct antiviral efficacy of interferon alpha (IFN-α) treatment on hepatic hepatitis B virus (HBV) replication has not been well defined. Using HBV-infected humanized uPA/SCID mice we recently showed that treatment with conventional IFN-α induced short-lived (< 24 hours) suppression of HBV replication and the responsiveness of infected human hepatocytes to IFN-α was impaired (Gastroenterology 2011:140;2074-83). Aim of this study was to investigate whether the use of pegylated interferon-α (peg-IFN-α), alone or in combination with entecavir (ETV), could induce more effective antiviral control in HBV-infected mice and improve hepatocyte responsiveness to IFN-α. Methods: HBV-infected humanized uPA/SCID mice displaying median viral titers of 3.6x10E8 HBV-DNA/ml (n=23) received either peg-IFN-α monotherapy (25ng/gr, twice/week; n=6), ETV monotherapy (100ng/gr/day; n=6), or both drugs in combination (n=6), while 5 animals were left untreated as controls. Serological changes were determined prior to, and after 2 and 4 weeks of treatment. Intrahepatic analyses were performed by qRT-PCR and immunohistochemistry. Results: After 4 weeks of therapy, the median reduction in viremia was 1.4Log, 3.4Log and 3.3Log in mice treated with peg-IFN-α, ETV or the combination thereof, respectively. Despite the larger viremia reduction provided by ETV, the reduction of intrahepatic HBcAg expression levels was more pronounced in mice receiving peg-IFN-α alone or combination therapy, than in mice receiving ETV monotherapy. Peg-IFN-α alone or in combination also reduced the levels of circulating HBsAg (0.21Log; 0.5Log) and HBeAg (0.55Log; 0.66Log), whereas the median decrease in these viral antigens was less pronounced in mice receiving ETV monotherapy (HBsAg=Δ0.13Log; HBeAg=Δ0.24Log). Remarkably, the progressive decrease of viral antigens was accompanied by an improved responsiveness to IFN-α, which was demonstrated by the ability to induce STAT translocation and enhancement of human interferon stimulated genes (ISG) in mice continuously treated with peg-IFN-α. Conclusions: The strong reduction of intrahepatic viral loads and antigenemia achieved in vivo in the absence of adaptive immune responses emphasizes the qualitatively different antiviral effects induced by peg-IFN-α compared to polymerase inhibitors, thus providing a rationale for exploring the potential of therapeutic strategies based on the combination of distinct types of antiviral agents.

StephenW

标题聚乙二醇α-干扰素单独或联合恩替卡韦恢复的ISG反应能力和降低肝内的病毒载量与抗原在乙肝病毒感染人性化的小鼠
主讲人：莉娜Allweiss
作者：属Allweiss1 *，M.Lütgehetmann1，2，T. Volz1，T. Bornscheuer1，AW lohse1，MA4研究Petersen3，H. K. Klumpp4，与Fletcher4，Dandri1研究
所属：1Internal医药，微生物学，病毒学及卫生2Medical大学医学中心的汉堡Eppendorf公司，3Liver中心，圣乔治Asklepiosklinik，汉堡，德国，4Hoffmann-La Roche公司，纳特利，新泽西州，美国。 * [email protected]
背景：α-干扰素（α-干扰素）治疗肝B型肝炎病毒（HBV）复制的直接抗病毒疗效一直没有得到很好的定义。使用人性化的UPA / SCID小鼠HBV感染，我们最近发现，与传统的α-干扰素诱导的短暂（<24小时）抑制乙肝病毒复制和感染人类肝细胞的反应，α-干扰素治​​疗受损（胃肠病学2011:140 ; 2074-83）。本研究的目的是调查是否使用聚乙二醇α-干扰素（PEG-IFN-α），单独或联合恩替卡韦（ETV），可诱导乙肝病毒感染的小鼠更有效的抗病毒药物控制和改善肝细胞​​干扰素的反应α-。
方法：HBV感染的人性化UPA / SCID小鼠显示的3.6x10E8 HBV-DNA/ml（N = 23）的平均病毒滴度接受PEG-α-干扰素单一（25ng/gr，两次/周; N = 6），ETV单一（100ng/gr/day，N = 6），或都在结合药物组（n = 6），而5只留下控制未经处理的。血清学变化进行了测定前，后2和4周的治疗。肝内进行qRT-PCR和免疫组化分析。
结果：经过4周的治疗，在病毒血症的中位数减少了1.4Log，3.4Log和在小鼠PEG-IFN-α，ETV或组合治疗3.3Log，分别。尽管ETV提供了更大的病毒血症减少，肝内HBcAg表达水平的降低更明显比接受ETV单药治疗的老鼠在接受PEG-IFN-α单独或联合治疗小鼠。 PEG-IFN-α单独或合并也降低循环HBsAg的（0.21Log; 0.5Log）的水平和HBeAg（0.55Log; 0.66Log），而在这些病毒抗原的中位数下降是在接受ETV单药治疗的老鼠少（ =Δ0.13Log乙肝表面抗原，e抗原=Δ0.24Log）。值得注意的是，病毒抗原的逐步减少，伴随着α-干扰素，证明这是由STAT易位和提高人类在不断与PEG-α-干扰素治​​疗的小鼠干扰素刺激基因（ISG）的能力，促使改进的响应。
结论：肝内体内的病毒载量和适应性免疫反应的情况下取得的抗原的强还原性，强调聚合酶抑制剂相比，PEG-IFN-α诱导的定性不同的抗病毒效果，从而提供了一种潜在的治疗策略探索的理由不同类型的抗病毒药物相结合的基础上。