Hydrodynamic injection of viral DNA: A mouse model of acute hepatitis B virus infection
Hydrodynamic injection of viral DNA: A mouse model of acute hepatitis B virus infection
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DOI：
10.1073/pnas.202398599
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发表时间：
2002-10-15
影响因子：
11.1
通讯作者：
Chisari, FV
中科院分区：
文献类型：
--
作者：
Yang, PL;Althage, A;Chisari, FV
B型肝炎病毒（HBV）是肝脏特异性病原体的原型，其中免疫系统未能建立有效的应答导致慢性感染。我们对H8 V的免疫反应的理解是不完整的，主要是由于这种病原体的宿主限制和现有实验模型的局限性。我们已经开发了一种小鼠模型，用于研究人类HBV复制、免疫原性和控制。在体内用具有复制能力的、超长的、线性的HBV基因组转染肝细胞后，合成病毒抗原和复制中间体，并将病毒分泌到血液中。病毒抗原早在转染后7天就从血液中消失，与抗病毒抗体的出现一致。在抗病毒CD 8 + T细胞出现后，HBV转录本和复制中间体在第15天从肝脏中消失。相比之下，病毒在NOD/Scid小鼠转染后持续至少81天，NOD/Scid小鼠缺乏功能性T细胞、B细胞和自然杀伤（NK）细胞。因此，HBV的流体动力学转染的结果取决于宿主免疫应答，就像在自然感染期间一样。我们所描述的方法将允许在严格控制的体内系统中检查病毒动力学，应用诱变方法研究体内HBV生命周期，以及使用免疫调节和免疫效应功能已被删除或过表达的转基因小鼠解剖对HBV的免疫应答。此外，该方法为研究其他已知和待发现的肝脏特异性病原体提供了原型。
Hepatitis B virus (HBV) is a prototype for liver-specific pathogens in which the failure of the immune system to mount an effective response leads to chronic infection. Our understanding of the immune response to H8V is incomplete, largely due to the narrow host restriction of this pathogen and the limitations of existing experimental models. We have developed a murine model for studying human HBV replication, immunogenicity, and control. After transfection of hepatocytes in vivo with a replication-competent, over-length, linear HBV genome, viral antigens and replicative intermediates were synthesized and virus was secreted into the blood. Viral antigens disappeared from the blood as early as 7 days after transfection, coincident with the appearance of antiviral antibodies. HBV transcripts and replicative intermediates disappeared from the liver by day 15, after the appearance of antiviral CD8 + T cells. In contrast, the virus persisted for at least 81 days after transfection of NOD/Scid mice, which lack functional T cells, B cells, and natural killer (NK) cells. Thus, the outcome of hydrodynamic transfection of HBV depends on the host immune response, as it is during a natural infection. The methods we describe will allow the examination of viral dynamics in a tightly controlled in vivo system, the application of mutagenesis methods to the study of the HBV life cycle in vivo, and the dissection of the immune response to HBV using genetically modified mice whose immunoregulatory and immune effector functions have been deleted or overexpressed. In addition, this methodology represents a prototype for the study of other known and to-be-discovered liver-specific pathogens.