APASL2013 缺乏HBsAg的转基因小鼠缺明显诱导干扰素的反应

StephenW

Abstract

The Interferon Response Is Significantly Induced by Hepatitis B Virus Replication in HBV-transgenic Mice Lacking the HBsAg
     
Catherine Isabell Real1, Ruth Broering1, Martin Trippler1, Kerstin Jahn-Hofmann2, Ludger Ickenstein2, Matthias John2, Kathrin Kleinehr1, Sabrina Driftmann1, Thekla Kemper3, Hans-Peter Vornlocher4, Reinhold Schirmbeck5, Mengji Lu3, Guido Gerken1, Joerg Friedrich Schlaak1
1Clinic of Gastroenterology and Hepatology, University Hospital Essen, Essen, 2Roche Kulmbach GmbH, Kulmbach, 3Dept. of Virology, University Hospital of Essen, Essen, 4Axolabs GmbH, Kulmbach, 5Dept. of Internal Medicine, University of Ulm, Ulm, Germany

Introduction: Previously, it has been suggested that chronicity of hepatitis B infection may be facilitated by an attenuation of innate and adaptive immune responses through HBsAg. Here, we have studied the immunological effects of HBV replication in a transgenic mouse model, lacking the HBsAg (tg1,4HBV-S-mut). We also characterized siRNA-mediated HBV suppression in this system.
Methods: Nanolipid-formulated siRNAs specific to the HBxAg mRNA, were injected intravenously into HBV-S-mut mice. HBV negative littermates were used as controls. After 48h and 10d tissue (liver, heart, spleen, kidney) was prepared and expression of HBV-RNA, IFN-β, ISG15, IFI-T1, IP-10, TGF-β, TNF-α, IL-6 and IL-10 was determined by qtRT-PCR. HBV DNA in liver tissue was determined by Southern blot. Then, transcriptome analysis was performed using total liver tissue. ELISAs and cytokine arrays were performed to detect HBeAg and cytokine levels in serum.
Results: Single injection of siRNAs against HBV led to suppression of HBV DNA after 48h which was sustained for more than 10 days. The HBeAg serum levels were also reduced about 90% after treatment with HBxAg-specific siRNA. The expression of IFN-β, IFI-T1 and ISG15 was up-regulated in HBV positive animals compared to control animals, which could be normalized by treatment with HBxAg-specific siRNAs. Transcriptome analysis after siRNA mediated knockdown exhibited different up-regulated and down-regulated genes in tgHBV-S-mut mice compared to littermates, which could be returned to basal expression after siRNA-mediated suppression of HBV. In tgHBV mice microRNA analysis identified regulated miRNAs, which are associated with HBV infection and progression of hepatocellular carcinoma.
Conclusions: In contrast to human liver from HBV patients, in HBV transgenic mouse model lacking HBsAg viral replication is associated with expression of IFNs and ISGs. Thus, we hypothesize that HBsAg is suppressing the IFN induction and ISG response in vivo. This hypothesis will be challenged in HBsAg transgenic mice.


Assigned speakers:
Mrs. Catherine Real, University Hospital Essen , Essen , Germany

Assigned in sessions:
07.06.2013, 08:30-17:30, PT-3, HEP B Basic, Exhibition Hall

StephenW

简介：此前，曾有人建议，B型肝炎感染的慢性化，可促进由先天和适应性免疫应答的衰减通过的HBsAg。在这里，我们在转基因小鼠模型研究HBV复制的免疫效果，缺乏乙肝表面抗原（TG1，4HBV-S-MUT）。特征还在于在该系统中的siRNA介导的HBV抑制。
方法：Nanolipid配制特定的HBxAg的mRNA的siRNA，HBV-S-静音小鼠静脉注射到。 HBV阴性作为对照窝。后48h和10D的组织（肝，心，脾，肾）编制及表达HBV-RNA，IFN-β，ISG15，IFI-T1，IP-10，TGF-β，TNF-α，IL-6和IL -10乃由qtRT-PCR。测定肝组织中HBV DNA印迹。然后，进行总肝组织转录组分析。 ELISA试剂盒及细胞因子的阵列进行检测HBeAg和血清中细胞因子水平。
结果：单注射抗HBV的siRNA抑制HBV DNA，这是持续了10多天的48小时后。与HBxAg特异性siRNA治疗后HBeAg的血清水平也减少了约90％。 IFN-β的表达，IFI-T1和ISG15的上调，这可能是归一化通过与HBxAg特定的siRNA处理的对照组动物相比，在​​HBV阳性动物。 siRNA介导的转录组分析后击倒在tgHBV-S-MUT小鼠表现出不同的上调和下调基因相比，窝，这可能是后返回到基础表达的siRNA介导的抑制HBV。在tgHBV小鼠microRNA分析确定调节的miRNA，这都与HBV感染与肝癌的进展。
结论：相反从HBV的患者，在HBV的转基因小鼠模型缺乏HBsAg的病毒复制，对人体肝与表达干扰素和ISGs。因此，我们推测，乙肝表面抗原是抑制体内的干扰素诱导和ISG响应。这一假说将受到挑战HBsAg转基因小鼠。

MP4

结论能否解释下
表面抗原抑制免疫?
还是?

StephenW

回复 MP4 的帖子

是的.

肝肠欲断

都看不懂