IL-21-based therapies induce clearance of hepatitis B virus persistence in mouse models
IL-21-based therapies induce clearance of hepatitis B virus persistence in mouse models
复制标题
基于 IL-21 的疗法可诱导小鼠模型中持久存在的乙型肝炎病毒的清除
DOI：
10.7150/thno.35331
复制
发表时间：
2019-01-01
期刊：
影响因子：
12.4
通讯作者：
Xie，Youhua
中科院分区：
文献类型：
--
作者：
Shen，Zhongliang;Liu，Jing;Xie，Youhua
慢性乙肝病毒感染会导致肝炎、肝硬变和肝细胞癌。共价闭合环状DNA(CccDNA)是唯一的病毒转录模板，不受当前治疗方案的影响，构成了乙肝病毒持久性的关键决定因素。需要对cccDNA具有明显疗效的新疗法。方法：在此之前，我们使用来自临床分离株(称为BPS)的复制子质粒建立了一种乙肝病毒持续存在的小鼠模型，并确认IL-21是一种有效的清除诱导剂。我们还描述了另一种基于体内产生的模拟cccDNA的持久化小鼠模型，称为重组ccccDNA(RcccDNA)。在这项工作中，使用这些模型测试了基于IL-21的基因和细胞疗法的有效性。结果：在两种模型中，一次注射表达小鼠IL-21的腺相关病毒(AAV)可以高效地清除血清中的两种乙肝标志物，更重要的是清除小鼠肝脏中的BPS DNA和rcccDNA。从机制上讲，IL-21诱导的清除与CD8+T细胞的活化和肝脏的浸润有关，CD8抗体注射对AAV-IL-21的疗效有负面影响。更值得注意的是，过继转移来自AAV-IL-21治愈小鼠的CD8+T细胞在受体HBV持久型小鼠中产生了清除。此外，治愈的小鼠具有长期的记忆力，可以防止再次受到攻击。最重要的是，输注用IL-21蛋白和乙肝病毒抗原体外刺激的治疗初治小鼠的脾细胞也能诱导治疗初治小鼠的清除。结论：以IL-21为基础的基因和细胞疗法是治疗慢性乙肝的有效候选方法，可通过激活CD8+T细胞清除携带cccDNA的肝细胞，并伴有长期记忆保护作用。
Chronic hepatitis B virus (HBV) infection causes hepatitis, liver cirrhosis and hepatocellular carcinoma. Covalently closed circular DNA (cccDNA) is the sole viral transcription template and not affected by current treatment options, constituting a key determinant of HBV persistence. Novel therapeutics with demonstrable effectiveness against cccDNA are required. Methods: Previously, we established an HBV persistence mouse model using replicon plasmid derived from a clinical isolate (termed BPS) and identified IL-21 as a potent clearance-inducer. We also described another persistence mouse model based on cccDNA mimics produced in vivo termed recombinant cccDNA (rcccDNA). In this work, effectiveness of IL-21-based gene and cellular therapies was tested using these models. Results: In both models of HBV persistence, single injections with adeno-associated virus (AAV) expressing murine IL-21 highly efficiently induced clearance of both HBV markers from serum, and more importantly, BPS DNA and rcccDNA from mouse liver. Mechanistically, IL-21-induced clearance was associated with activation and liver infiltration of CD8+ T cells, and CD8 antibody injections negatively affected AAV-IL-21 effectiveness. More notably, adoptive transfer of CD8+ T cells from AAV-IL-21-cured mice engendered clearance in acceptor HBV persistence mice. Furthermore, cured mice were protected against re-challenge with long-lived memory. Most significantly, infusion of splenocytes from treatment-naïve mice stimulated ex vivo with IL-21 protein and HBV antigen could also induce clearance in treatment-naïve mice. Conclusion: These data demonstrate IL-21-based gene and cellular therapies as valid candidates for treating chronic HBV infections, with potential in removing cccDNA-harboring hepatocytes via activated CD8+ T cells accompanied by long-term protective memory.