Abstract
The use of replication-competent hepatitis B virus (HBV) DNA to construct a mouse model will help explore antiviral treatment strategies for more than 240 million patients infected with HBV worldwide. Eradication of chronic HBV infection can effectively block the adverse consequences of HBV-induced hepatic cirrhosis, failure and carcinoma. The core reason that HBV is difficult to eradicate is that most of infected people develop chronic HBV infection due to the establishment of immune tolerance. Here, we introduce a mouse model of adeno-associated virus (AAV)-HBV transfection, which produces HBV surface antigen (HBsAg) that can be maintained for more than 6 months. During virus replication, intermediates, transcripts, and proteins can be detected in peripheral blood. At the same time, the prerequisite for studying liver disease formation and immunotherapy through in vitro experiments is to isolate hepatic subgroup cells. Here, we describe a cell sorting method based on liberase perfusion technology combined with low-speed centrifugation and magnetic bead antibody labeling to purify hepatic parenchymal cells (PCs) and non-parenchymal cells (NPCs) step by step from murine liver, such as hepatic sinusoidal endothelial cells (LSECs) and Kupffer cells (KCs), which will help accelerate the study of the genetic and clearance mechanistic of chronic HBV infection.
Highlights
Hepatitis B virus (HBV) has an extremely narrow host-range, such as humans [1], chimpanzees [2], Mauritian cynomolgus monkey [3], treeshrew [4, 5] and woodchuck [6, 7]
Studies on the HBV mouse models have gone through several generations, involved in HBV DNA transgenic mice [9], HBV DNA transfected mice [10], HBV rc-cccDNA transfected mice [11], HBV-infected liver chimeric humanized mice and CRISPR/Cas9 technological NRG/FahÀ/À immune deficient mice [12]
We will introduce how to establish this mouse model, the detectable HBV surface antigen (HBsAg) protein persists in blood or liver more than 6 months and has been widely used as chronic infection model [19]
Summary
Hepatitis B virus (HBV) has an extremely narrow host-range, such as humans [1], chimpanzees [2], Mauritian cynomolgus monkey [3], treeshrew [4, 5] and woodchuck [6, 7]. Studies on the HBV mouse models have gone through several generations, involved in HBV DNA transgenic mice [9], HBV DNA transfected mice [10], HBV rc-cccDNA transfected mice [11], HBV-infected liver chimeric humanized mice and CRISPR/Cas technological NRG/FahÀ/À immune deficient mice [12]. Their advantages and disadvantages are as follows, separately: (1) 1.3-fold HBV genome transgenic mice support the expression of viral RNA and viral proteins in the liver, and can develop complete pgRNA, viral assembly and viral secretion during in the viral cycle, and support endosomal antiviral [13]. Different from immunodeficient mice such as liver humanized mice, the mice can express HBV Ags, and the immune system can recognize the Ags to stimulate virus-specific immune responses This model system can be used to test HBV cure strategies and study HBV immunology [15, 16]. The specific mechanism will be verified using isolated mouse hepatocytes and the corresponding mechanism will be explained
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