AASLD2014:乙肝病毒感染人体肝细胞核苷酸类似物提高干扰素的

StephenW

1740
Nucleotide analogue improves interferon responsiveness in HBV-infected human hepatocytes
Masataka Tsuge1,3, Nobuhiko Hiraga1,2, Eisuke Murakami1,2, Michio Imamura1,2, Hiromi Abe1,2, Daiki Miki1,2, Hidenori Ochi1,2, C. Nelson Hayes1,2, Kazuaki Chayama1,2;
1Department of Gastroenterology and Metabolism, Hiroshima university, Hiroshima, Japan; 2Liver Research Project Center, Hiroshima university, Hiroshima, Japan; 3Natural Science Center for Basic Research and Development, Hiroshima university, Hiroshima, Japan

Background: It has been reported that interferon treatment could reduce HBs antigen (HBsAg) production in patients with chronic hepatitis B virus (HBV) infection. However, only limited HBsAg reduction is observed in patients treated with interferon therapy. One cause of this limitation may be that interferon responsiveness in human hepatocytes is suppressed by HBV infection, and, therefore, interferon stimulated genes (ISGs) are not induced sufficiently to promote anti-viral effects. In the present study, we analyzed whether the suppression of HBV replication using nucleotide analogues (NAs) could improve interferon responsiveness in HBV infected human hepatocytes. Methods: Thirty-seven chronic hepatitis B patients were enrolled. Twenty patients underwent sequential interferon therapy, which included 6 months of conventional interferon therapy, running from one month prior to discontinuation until 5 months after discontinuation of NA therapy. The remaining 17 patients underwent interferon mono-therapy. Serum HBsAg titers were measured every year for 5 years after interferon therapy. To confirm the clinical results, we performed an in vitro study using T23 cells, which were generated from HepG2 cells stably transfected with an HBV expression plasmid. T23 cells were treated with or without the NA entecavir for 5 days. The cells were then treated with IFN for 6 hours and harvested. To evaluate the interferon responsiveness, ISG mRNA levels were quantified by real time PCR. Results: In the clinical study, more than 1 Log IU/ml reduction of HBsAg titer was achieved in 11 of 37 patients (interferon mono-therapy: 2, Sequential therapy: 9). By univariate analysis, the following factors, gender, serum HBsAg level, the existence of HBeAg, and prior NA therapy, were associated with HBsAg reduction (P=0.007, P=0.027, P=0.031, P=0.037, respectively). From the clinical results, it was predicted that interferon responsiveness might be improved by prior NA therapy. To verify these results, in vitro experiments were performed. In the absence of HBV, the ISGs MxA and OAS1 were significantly induced by interferon treatment (19.2-fold, 9.7-fold, respectively). However, in T23 cells, inductions of these ISGs was suppressed (P=0.0495, P=0.0495, respectively). After entecavir treatment, interferon responsiveness was restored and ISG induction increased (P=0.0495, P=0.0339, respectively). Conclusions: Prior NA therapy could improve interferon responsiveness in HBV infected human hepatocytes. To improve the anti-viral effects in chronic hepatitis B patients, it might be necessary to revise the way of using NAs and interferons.

StephenW

1740
乙肝病毒感染人体肝细胞核苷酸类似物提高干扰素的反应
正隆Tsuge1,3，信彦Hiraga1,2，英介Murakami1,2，道夫Imamura1,2，博美Abe1,2，大树Miki1,2，秀典Ochi1,2，C.尼尔森Hayes1,2，一明Chayama1,2;
教研室胃肠病学和代谢，广岛大学，广岛，日本; 2Liver研究项目中心，广岛大学，广岛，日本; 3Natural科学中心的基础研究和开发，广岛大学，日本广岛

背景：据报道，干扰素治疗可以减少在慢性乙型肝炎病毒（HBV）感染HBs抗体抗原（HBsAg）的生产。然而，只有有限的HBsAg还原在用干扰素疗法治疗的患者中观察到。此限制的一个原因可能是，干扰素的反应在人肝细胞中由HBV感染抑制，并且，因此，干扰素刺激基因（的ISG）不会引起足以促进抗病毒的效果。在本研究中，我们分析了HBV的复制使用的核苷酸类似物（NAS）的抑制是否可以改善在HBV感染的人肝细胞中的干扰素应答。方法：三十七名慢性乙肝患者入组。 20例患者接受干扰素序贯治疗，其中包括6个月常规干扰素治疗，从一个月前终止运行，直到NA治疗停药后5个月。其余17例患者接受干扰素单一治疗。血清HBsAg滴度测定每年都为干扰素治疗后的5年。确认临床效果，我们进行了体外用T23细胞中，从HepG2细胞稳定转染了HBV表达质粒产生的研究。 T23细胞用或不用NA恩替卡韦，连续5天。然后将细胞用IFN处理后6小时，收获。为了评估干扰素反应，ISG表达水平进行实时PCR定量。结果：在临床研究中，超过1登录IU减少HBsAg的滴度/毫升中1137的患者达到（干扰素单一疗法：2，序贯疗法：9）。单因素分析显示，以下几个因素，性别，血清HBsAg水平，HBeAg的存在，和以前的NA治疗，均与乙肝表面抗原降低（P = 0.007，P =0.027，P =0.031，P=0.037）。从临床结果可以预测，干扰素响应可能由现有NA治疗得到改善。为了验证这些结果，在体外实验。在没有乙肝，该的ISG的MxA和OAS1干扰素治疗（19.2倍，9.7倍，分别）被显著诱导。然而，在T23细胞中，这些的ISG导入装置被抑制（P =0.0495，P =0.0495，分别）。恩替卡韦治疗后，干扰素反应性恢复和ISG诱导增加（P=0.0495，P=0.0339，分别）。结论：此前NA治疗能改善乙肝病毒感染的人类肝细胞干扰素反应。以提高在慢性乙型肝炎患者的抗病毒效果，它可能有必要修改使用NAS和干扰素的方法。

lgs1

这个是先干扰还是先核甘啊？

StephenW

回复 lgs1 的帖子

先核甘.

lgs1

中国好像也在开展类似临床实验