用于乙型肝炎病毒相关研究的人源化嵌合肝脏小鼠模型应用进展

人源化嵌合肝脏小鼠模型是一类具有肝损伤及免疫缺陷特点的小鼠模型。该小鼠模型体内携带人肝细胞，成功攻克了由于乙型肝炎病毒（HBV）宿主范围狭窄导致的HBV易感动物模型缺乏问题。近年来，随着各种人源化嵌合肝脏小鼠模型的逐渐完善，使得HBV相关动物研究的顺利开展成为可能。从Alb-uPA嵌合肝脏小鼠模型、Alb-rtTA/TRE-uPA嵌合肝脏小鼠模型、FRG嵌合小鼠模型到TK-NOG嵌合小鼠模型、人源化双嵌合小鼠模型逐渐克服了宿主物种限制和疾病建模方面的障碍。人源化双嵌合小鼠模型同时具备人免疫系统及人肝细胞，在HBV免疫机制研究及疫苗研发中具有极大的应用前景。了解不同人源化嵌合肝脏小鼠模型的制作情况、适用情况及优缺点，可为相关研究中HBV易感动物模型的选择提供参考。     

免疫调节剂在慢性乙型肝炎病毒感染治疗中的作用

慢性乙型肝炎病毒（HBV）感染是亚洲人健康的一个主要威胁。为了设计一个更好的治疗方案，必须明白以下关于HBV病毒残留的机制。免疫学研究指出HBV病毒的特异性T细胞应答削弱是导致慢性感染的主要原因。然而对HBV的多种特殊、强力的T细胞应答与长时期控制而不是消灭病毒有关。此外，在异源造血细胞移植中血清乙型肝炎病毒表面抗原（HB-sAg）的清除，HBsAg血清转化与捐赠者乙型肝炎病毒核心特性抗原CIM＋T淋巴细胞数激活有关。     

肝炎病毒对造血干细胞移植的影响

目的:探讨肝炎病毒对造血干细胞移植(HSCT)的影响。方法:回顾性分析我院21例肝炎病毒感染接受HSCT治疗的患者的临床资料。乙型肝炎病毒(HBV)感染18例,其中供受者同时有HBV感染4例,供者有HBV感染3例,受者有HBV感染11例,供者、受者移植前后服用拉米夫定预防和治疗HBV感染。丙型肝炎病毒(HCV)感染3例,均为受者感染,移植免疫恢复后给予α干扰素治疗。结果:HSCT后植活平均15(9～31)d。HSCT后发生急性移植物抗宿主病(GVHD)9例(47%),发生慢性GVHD7例(37%)。HSCT后无肝静脉闭塞症发生。HBV感染供者,HSCT后无爆发性乙型病毒性肝炎发生。1例(1/18)HBV感染患者HSCT后6个月停用拉米夫定而发生爆发性乙型病毒性肝炎。1例(1/18)受者HBsAg HBeAg HBcAb(+),供者HBsAb(+),HSCT3个月后患者HBsAb HBeAb HBcAb(+),HBV-DNA阴性。3例HCV感染患者HSCT后用α干扰素治疗,2例HCV-RNA转阴。结论:HBV感染的供受者,在行HSCT过程中,应用拉米夫定能够有效预防移植后乙型病毒性肝炎的复燃,丙型病毒性肝炎的患者应用干扰素治疗可使HCV-RNA转阴。初步认为,肝炎病毒感染可能并非是HSCT的禁忌症。     

重型病毒性肝炎病原学特点及转归

探讨重型病毒尾肝炎的病原学特点。收集各型重型病毒性肝炎418例，分析其病原学分型及乙型肝炎病毒不同病原学模式与重型肝炎预后的关系。急性重型肝炎以甲型、戊型及乙型病毒性肝炎为主，乙型肝炎病毒感染治愈后病毒阴转率较高。亚急性有慢性重型肝炎以乙型肝炎病毒毒感染居首，占92．8％。在乙型肝炎病毒感染的病原学模式中，以HBsAgHBeAbHBcAb阳性的重型肝炎发病及死亡率最高。乙型肝炎病毒与其他肝炎病毒重叠感染与单独感染比较，死亡率无显著差异。单纯TTV感染可导致重型肝炎。重型肝炎发病后HBVDNA可自然阴转，阴转率可达53．6％。重型肝炎仍以乙型肝炎病毒病毒感染为主。乙型肝炎病毒前C区发生基因突变可能较易发生重型肝炎。     

人鼠嵌合肝小鼠模型的建立及在HBV致病研究中的应用

病毒性肝炎一直是全球范围内一个沉重的肝病负担,其中以乙型肝炎最为严重。由于HBV仅感染灵长类动物,因此其所致肝病很难在动物模型中被复制。人鼠嵌合肝小鼠模型是指将人源肝细胞移植入免疫缺陷的肝损伤小鼠肝内而形成的嵌合体小鼠。由于小鼠肝内嵌合有人肝细胞,可以在其体内模拟HBV的感染,利于进行进一步的机制研究。目前该模型主要以白蛋白启动子调控尿激酶(Alb-uPA)小鼠模型和延胡索酰乙酰乙酸水解酶(Fah~(-/-))小鼠模型为基础,在人源肝组织和外周血中均可检测到HBV,并已应用于HBV突变、HBV基因功能、抗病毒药物效果以及HBV与免疫系统的相互作用等方面的研究。本文就传统HBV感染动物模型的利弊、人鼠嵌合肝小鼠模型的研究进展和改进方法进行综述。     

肝炎病毒基因工程抗体的研究

0引言乙型病毒性肝炎?丙型病毒性肝炎分别是由乙型肝炎病毒(HBV)和丙型肝炎病毒(HCV)感染引起的世界性传染病[1-10].HBV?HCV感染不仅引起急性?慢性病毒性肝炎,而且与肝硬化和肝细胞癌的发生和发展密切相关[11-20].流行病学调查表明,HB V感染的阳性率为10%,抗-HCV的阳性率为3.2%,对我国人民健康危害极大.因此,急需研究有效的针对乙型?丙型病毒性肝炎的防治措施.其中肝炎病毒人源化基因工程抗体及其应用是目前的主要研究方向之一.1乙型肝炎病毒?丙型肝炎病毒基因工程抗体的研究背景目前在我国引起慢性病毒性肝炎的肝炎病毒类型主要是H…     

人鼠嵌合肝模型的建立与鉴定

动物模型在肝病学的发展中起重要作用。近几年，在了解特异基因功能、代谢通路和疾病进程方面，啮齿类动物模型发挥了巨大作用。人鼠嵌合肝是利用人肝细胞异种移植到受体鼠肝内建立的新型动物模型。受体鼠可分为免疫缺陷小鼠或诱导免疫耐受大鼠^[1,2]，两者均能初步建立HBV感染的人鼠嵌合肝动物模型。本文主要介绍人鼠嵌合肝相关内容，包括移植细胞来源、建立途径、     

人La自身抗原蛋白与肝炎病毒的关系

0 引言乙型和丙型肝炎呈全球分布,目前全世界有1.7亿人感染丙型肝炎病毒(HCV),3.5亿人感染乙型肝炎病毒(HBV),其中相当一部分感染者发展为慢性肝炎,少数还发展成肝硬化甚至肝癌,对人类健康造成极大危害,至今缺乏有效的控制.肝炎病毒在肝细胞中的长期存在,复制扩散是肝炎病毒慢性感染的关键一步,这两种病毒在体内感染的分子生物学机制一直是研究者们关注的重点,许多的研究越来越显示 La 自身抗原蛋白与其关系密切,本文简要介绍近年来在这方面的研究进展.     

CD4＋ CD25＋调节性T细胞与乙型肝炎病毒感染的研究进展

CD4＋ D25＋ 调节性T细胞（Treg）是近年来发现的一群具有免疫抑制功能的细胞亚群,其独特的作用方式及功能特征使其在自身免疫性疾病、移植免疫、肿瘤免疫和感染免疫中发挥着重要作用.近年的研究表明,CD4＋ CD25＋ Treg与乙型肝炎病毒（HBV）感染后耐受、发病和转归相关.     

戊型肝炎

大庆地区戊型肝炎病毒亚临床感染情况调查，戊型肝炎病毒嵌合蛋白的免疫应答的初步研究，     

Animal Models of Hepatitis B Virus Infection–Success,Challenges, and Future Directions

Chronic hepatitis B virus (HBV) infection affects more than 250 million people worldwide, which greatly increases the risk for terminal liver diseases, such as liver cirrhosis and hepatocellular carcinoma (HCC). Even though current approved antiviral therapies, including pegylated type I interferon (IFN) and nucleos(t)ide analogs, can effectively suppress viremia, HBV infection is rarely cured. Since HBV exhibits a narrow species tropism and robustly infects only humans and higher primates, progress in HBV research and preclinical testing of antiviral drugs has been hampered by the scarcity of suitable animal models. Fortunately, a series of surrogate animal models have been developed for the study of HBV. An increased understanding of the barriers towards interspecies transmission has aided in the development of human chimeric mice and has greatly paved the way for HBV research in vivo, and for evaluating potential therapies of chronic hepatitis B. In this review, we summarize the currently available animal models for research of HBV and HBV-related hepadnaviruses, and we discuss challenges and future directions for improvement.     

Animal model for study of human hepatitis viruses

Human hepatitis B virus (HBV) and hepatitis C virus (HCV) infect only chimpanzees and humans. Analysis of both viruses has long been hampered by the absence of a small animal model. The recent development of human hepatocyte chimeric mice has enabled us to carry out studies on viral replication and cellular changes induced by replication of human hepatitis viruses. Various therapeutic agents have also been tested using this model. In the present review, we summarize published studies using chimeric mice and discuss the merits and shortcomings of this model.     

Humanized chimeric uPA mouse model for the study of hepatitis B and D virus interactions and preclinical drug evaluation

No specific drugs are currently available against hepatitis delta virus (HDV), a defective virus leading to the most severe form of chronic viral hepatitis in man. The lack of convenient HDV infection models has hampered the development of effective therapeutics. In this study, na?ve and hepatitis B virus (HBV) chronically infected humanized uPA/SCID mice were employed to establish a small animal model of HBV/HDV coinfection and superinfection. For preclinical antiviral drug evaluation, the GMP version of the myristoylated preS-peptide (Myrcludex-B), a lipopeptide derived from the pre-S1 domain of the HBV envelope, was applied to prevent de novo HBV/HDV coinfection in vivo. Virological parameters were determined at serological and intrahepatic level both by real-time polymerase chain reaction (PCR) and by immunohistochemistry. Establishment of HDV infection was highly efficient in both HBV-infected and na?ve chimeric mice with HDV titers rising up to 1 × 10E9 copies/mL. Notably, HDV superinfection led to a median 0.6log reduction of HBV viremia, which although not statistically significant suggests that HDV may hinder HBV replication. In the setting of HBV/HDV simultaneous infection, a majority of human hepatocytes stained HDAg-positive long before HBV spreading was completed, confirming that HDV can replicate intrahepatically also in the absence of HBV infection. Furthermore, the increase of HBV viremia and intrahepatic cccDNA loads was significantly slower than in HBV mono-infected mice. Treatment with the HBV entry inhibitor Myrcludex-B, efficiently hindered the establishment of HDV infection in vivo. CONCLUSION: We established an efficient model of HBV/HDV infection to exploit mechanisms of viral interference in human hepatocytes and to test the efficacy of an HDV-entry inhibitor in vivo.     

HBV感染小鼠模型的研究进展

HBV由于其较强的种属特异性,目前尚缺乏理想的实验动物模型来阐明其具体的发病机制。目前有关HBV感染模型的研究大多集中在小鼠方面,并已取得了很大进展。综述了HBV转基因小鼠、HBV转染小鼠和人鼠嵌合肝脏HBV小鼠等模型的优缺点,提出合理使用这些模型有助于更好地阐明HBV的致病机制。     

高压水动力法建立表面抗原表达缺失的乙型肝炎转染小鼠模型及初步评估

目的探讨HBsAg在小鼠体内模型中对宿主免疫应答调节及病毒感染慢性化中的作用。方法在已有高压水动力法转染pAAV-HBV1.2质粒建立乙肝小鼠模型的基础上,通过点突变技术引入终止密码子至pAAV-HBV1.2质粒上的HBsAg读码框,构建HBsAg无法转录的突变质粒,再利用该质粒构建HBsAg表达缺失的乙肝小鼠模型。通过比较原始质粒建立的乙肝模型小鼠,检测分析外周血中HBV抗原与DNA等病毒感染指标,比较肝脏病变和免疫细胞浸润情况。结果成功构建了HBsAg缺失的乙肝模型小鼠:在乙肝模型小鼠和HBsAg缺失突变的模型小鼠外周血中均能检测到4 COI/mL水平的HBeAg,两组小鼠没有差异,但只有乙肝小鼠体内能检测到102 COI/mL以上的HBsAg和103至106拷贝/mL的HBV DNA,HBsAg缺失的小鼠中无法检测到HBsAg和HBV DNA;动态监测显示HBeAg在乙肝和HBsAg缺失突变的两组小鼠中的阳性率均逐步降低,在HBsAg缺失突变组中降低的更明显,第8周时仅有20%的小鼠为HBeAg阳性,而此时未突变乙肝组的阳性率为60%;免疫组织化学结果显示:在阴性对照小鼠的肝脏结构较完整,未见聚集的浸润免疫细胞,而乙肝和HBsAg缺失的乙肝小鼠肝小叶结构均松散,乙肝小鼠肝脏中有聚集的浸润性免疫细胞,HBsAg缺失的小鼠肝脏缺少免疫细胞的聚集,却存在严重的肝细胞毛玻璃样变性。结论成功构建了HBsAg缺失的乙肝转染小鼠模型;并发现HBsAg在HBV完整病毒颗粒包装和出胞,以及肝脏病变和免疫细胞浸润过程中发挥了重要的作用。     

Effects of hepatitis B virus infection on the interferon response in immunodeficient human hepatocyte chimeric mice

Complementary DNA microarray analysis of human livers cannot exclude the influence of the immunological response. In this study, complementary DNA microarray analysis was performed under immunodeficient conditions with human hepatocyte chimeric mice, and gene expression profiles were analyzed by hepatitis B virus (HBV) infection and/or interferon treatment. The expression levels of 183 of 525 genes upregulated by interferon treatment were significantly suppressed in response to HBV infection. Suppressed genes were statistically significantly associated with the interferon signaling pathway and pattern recognition receptors in the bacteria/virus recognition pathway (P = 1.0 × 10(-8) and P = 1.2 × 10(-8), respectively). HBV infection attenuated virus recognition and interferon response in hepatocytes, which facilitated HBV escape from innate immunity.     

Infection of human hepatocyte chimeric mouse with genetically engineered hepatitis B virus

Studies of hepatitis B virus (HBV) mutants have been hampered by the lack of a small animal model with long-term infection of cloned HBV. Using a mouse model in which liver cells were highly replaced with human hepatocytes that survived over a long time with mature human hepatocyte function, we performed transmission experiments of HBV. Human serum containing HBV and the virus produced in HepG2 cell lines that transiently or stably transfected with 1.4 genome length HBV DNA were inoculated. Genetically modified e-antigen-negative mutant strain also was produced and inoculated into the mouse model. A high-level (approximately 10(10) copies/mL) viremia was observed in mice inoculated with HBV-positive human serum samples. The level of viremia tended to be high in mice with a continuously high human hepatocyte replacement index. High levels and long-lasting viremia also were observed in mice injected with the in vitro generated HBV. The viremia continued up to 22 weeks until death or killing. Passage experiments showed that the serum of these mice contained infectious HBV. Genetically engineered hepatitis B e antigen-negative mutant clone also was shown to be infectious. Lamivudine effectively reduced the level of viremia in these infected mice. In conclusion, this mouse model of HBV infection is a useful tool for the study of HBV virology and evaluation of anti-HBV drugs. Our results indicate that HBeAg is dispensable for active viral production and transmission.     

Transfer of HBV genomes using low doses of adenovirus vectors leads to persistent infection in immune competent mice

Studies of mechanisms responsible for the persistence of hepatitis B virus (HBV) infection have been hindered by a lack of appropriate animal models. HBV genomes can be delivered to livers of mice using hydrodynamic injection or high doses of an adenoviral vector; these lead to clearance of HBV. We found that infection of immunocompetent mice with low doses of an adenoviral vector resulted in persistent HBV infection; the mice neither underwent seroconversion to production of antibodies against HBV nor developed a strong HBV-specific effector T-cell response. As in patients with chronic HBV infection, DNA vaccination failed to generate T cells that cleared infection. This model of persistent HBV infection could be used to study the pathogenesis of chronic HBV infection and develop new therapeutic strategies.