Differences between BALB/c and C57BL/6 Mice in Mouse Hepatitis Virus Replication in Primary Hepatocyte Culture (original) (raw)
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Immunobiology, 1992
The possible role of interferon-gamma (IFN-y) in the resistance of A/J mice to MHV3 infection was investigated. Monoclonal antibodies specific for IFN-y, CD4 and CDS molecules were administered in vivo to deplete selectively the IFN-y synthesized or the appropriate subset of T cells. The animals were then infected with MHV3 and the course of infection was followed by studying different parameters, such as, the mortality, the virus growth in the tissues and the IFN-y synthesis in sera and peritoneal exudates. After MHV3 infection, a full resistance of control A/J mice was observed, in contrast to the high mortality rate observed among the depleted animals, where higher virus titers were found in different tissues. The IFN-y synthesis in sera and peritoneal exudates of depleted mice, after MHV3 infection, drastically decreased when compared to that detected in control mice. The data presented are consistent with the hypothesis that IFN-y plays an essential role in the resistance of A/J mice to MHV3 infection.
Journal of Hepatology, 2009
Aim: Both hepatitis B virus (HBV) and hepatitis C virus (HCV) replicate in the liver and show resistance against innate immunity and interferon (IFN) treatment. Whether there is interference between these two viruses is still controversial. We investigated the interference between these two viruses and the mode of resistance against IFN. Methods: We performed infection experiments of either or both of the two hepatitis viruses in human hepatocyte chimeric mice. Huh7 cell lines with stable production of HBV were also established and transfected with HCV JFH1 clone. Mice and cell lines were treated with IFN. The viral levels in mice sera and culture supernatants and mRNA levels of IFN-stimulated genes were measured. Results: No apparent interference between the two viruses was seen in vivo. Only a small (0.3 log) reduction in serum HBV and a rapid reduction in HCV were observed after IFN treatment irrespective of infection with the other virus. In in vitro studies, no interference between the two viruses was observed. The effect of IFN on each virus was not affected by the presence of the other virus. IFN-induced reductions of viruses in culture supernatants were similar to those in in vivo study. Conclusion: No interference between the two hepatitis viruses exists in the liver in the absence of hepatitis. The mechanisms of IFN resistance of the two viruses target different areas of the IFN system.
Cytokine-Sensitive Replication of Hepatitis B Virus in Immortalized Mouse Hepatocyte Cultures
Journal of Virology, 2002
We have previously shown that alpha/beta interferon (IFN-␣/) and gamma interferon (IFN-␥) inhibit hepatitis B virus (HBV) replication by eliminating pregenomic RNA containing viral capsids from the hepatocyte. We have also shown that HBV-specific cytotoxic T lymphocytes that induce IFN-␥ and tumor necrosis factor alpha (TNF-␣) in the liver can inhibit HBV gene expression by destabilizing preformed viral mRNA. In order to further study the antiviral activity of IFN-␣/, IFN-␥, and TNF-␣ at the molecular level, we sought to reproduce these observations in an in vitro system. Accordingly, hepatocytes were derived from the livers of HBV-transgenic mice that also expressed the constitutively active cytoplasmic domain of the human hepatocyte growth factor receptor (c-Met). Here, we show that the resultant well-differentiated, continuous hepatocyte cell lines (HBV-Met) replicate HBV and that viral replication in these cells is efficiently controlled by IFN-␣/ or IFN-␥, which eliminate pregenomic RNA-containing capsids from the cells as they do in the liver. Furthermore, we demonstrate that IFN-␥, but not IFN-␣/, is capable of inhibiting HBV gene expression in this system, especially when it acts synergistically with TNF-␣. These cells should facilitate the analysis of the intracellular signaling pathways and effector mechanisms responsible for these antiviral effects.
Development of Mouse Hepatocyte Lines Permissive for Hepatitis C Virus (HCV)
PLoS ONE, 2011
The lack of a suitable small animal model for the analysis of hepatitis C virus (HCV) infection has hampered elucidation of the HCV life cycle and the development of both protective and therapeutic strategies against HCV infection. Human and mouse harbor a comparable system for antiviral type I interferon (IFN) induction and amplification, which regulates viral infection and replication. Using hepatocytes from knockout (ko) mice, we determined the critical step of the IFN-inducing/ amplification pathways regulating HCV replication in mouse. The results infer that interferon-beta promoter stimulator (IPS-1) or interferon A receptor (IFNAR) were a crucial barrier to HCV replication in mouse hepatocytes. Although both IFNARko and IPS-1ko hepatocytes showed a reduced induction of type I interferons in response to viral infection, only IPS-1-/-cells circumvented cell death from HCV cytopathic effect and significantly improved J6JFH1 replication, suggesting IPS-1 to be a key player regulating HCV replication in mouse hepatocytes. We then established mouse hepatocyte lines lacking IPS-1 or IFNAR through immortalization with SV40T antigen. Expression of human (h)CD81 on these hepatocyte lines rendered both lines HCVcc-permissive. We also found that the chimeric J6JFH1 construct, having the structure region from J6 isolate enhanced HCV replication in mouse hepatocytes rather than the full length original JFH1 construct, a new finding that suggests the possible role of the HCV structural region in HCV replication. This is the first report on the entry and replication of HCV infectious particles in mouse hepatocytes. These mouse hepatocyte lines will facilitate establishing a mouse HCV infection model with multifarious applications.
Mouse hepatitis virus (MHV) is a singular name for a group of murine coronaviruses that cause a wide spectrum of clinical outcomes ranging from subclinical infection to enteritis, hepatitis, and encephalitis. Enterotropic MHV strains, such as MHV-Y, initially replicate in epithelial cells of the gastrointestinal (GI) tract after infection of adult immunocompetent mice. Disease is acute and mild with minimal pathologic changes, and virus rarely disseminates to other organs (1-3). In contrast, morbidity and mortality are high in neonatal immunocompetent mice, and infection of immunocompromised mice can cause multisystemic, persistent infection with extended viral shedding (4) and high mortality (5). Polytropic MHV strains, such as MHV-A59 or MHV-JHM, initially replicate in the proximal respiratory tract epithelium after infection, then disseminate to many organs via viremia, lymphatic spread, or olfactory pathways from the nose to the brain. Infection of adult immunocompetent mice results in subclinical infection, hepatitis, and/or encephalitis (6), whereas infection of immunocompromised mice can cause severe disseminated disease with high mortality.
Journal of Hepatology, 1996
PLC/PRF/5 is a continuous human hepatocarcinoma cell line whose genome contains integrated HBV DNA and which secretes two of the hepatitis B virus envelope proteins (HBs and PreS2). This line is also susceptible to infection by hepatitis A virus and was therefore used to compare the effects of interferon on protein synthesis of these two viruses and to assess the interactions which occur between them during infection. Results showed that recombinant interferon alpha 2-a inhibited the expression of the two hepatitis B virus envelope antigens (HBs and PreS2) and of the only hepatitis A virus antigen in a dose-dependent fashion. Comparison of the effect of interferon on antigenic protein production of these two viruses, showed stronger inhibition of hepatitis A virus capsid antigen than of hepatitis B virus envelope antigens. Infection with hepatitis A virus also downregulates the expression of the two hepatitis B virus proteins. Considering the absence of cytotoxic effects from the doses used, this study confirms the relevance of this cellular model for the study of antiviral cytokines in vitro. It also provides a further rationale for the clinical evaluation of the therapeutic potential of interferons in severe hepatitis cases due either to hepatitis A virus alone or to superinfection of hepatitis B virus carriers by hepatitis A virus.
Non-cytolytic inhibition of hepatitis B virus replication in human hepatocytes
Journal of Hepatology, 2001
Background/Aims: Interferon-g (IFN-g) has been shown to abolish hepatitis B virus (HBV) gene expression and replication in HBV transgenic mice without destroying infected hepatocytes. We investigated the characteristics of IFN-g induced non-cytolytic inhibition of viral replication in human HBV infection.
Journal of Virology, 2006
Mouse hepatitis virus (MHV) does not induce interferon (IFN) production in fibroblasts or bone marrow-derived dendritic cells. In this report, we show that the essential IFN-β transcription factors NF-κB and IFN regulatory factor 3 are not activated for nuclear translocation and gene induction during infection. However, MHV was unable to inhibit the activation of these factors and subsequent IFN-β production induced by poly(I:C). Further, MHV infection did not inhibit IFN-β production mediated by known host pattern recognition receptors (PRRs) (RIG-I, Mda-5, and TLR3). These results are consistent with the notion that double-stranded RNA, produced during MHV infection, is not accessible to cellular PRRs.
PLoS ONE, 2013
Background & Aims: The interferon (IFN) system plays a critical role in innate antiviral response. We presume that targeted induction of IFN in human liver shows robust antiviral effects on hepatitis C virus (HCV) and hepatitis B virus (HBV). Methods: This study used chimeric mice harboring humanized livers and infected with HCV or HBV. This mouse model permitted simultaneous analysis of immune responses by human and mouse hepatocytes in the same liver and exploration of the mechanism of antiviral effect against these viruses. Targeted expression of IFN was induced by treating the animals with a complex comprising a hepatotropic cationic liposome and a synthetic double-stranded RNA analog, pIC (LIC-pIC). Viral replication, IFN gene expression, IFN protein production, and IFN antiviral activity were analyzed (for type I, II and III IFNs) in the livers and sera of these humanized chimeric mice. Results: Following treatment with LIC-pIC, the humanized livers of chimeric mice exhibited increased expression (at the mRNA and protein level) of human IFN-ls, resulting in strong antiviral effect on HBV and HCV. Similar increases were not seen for human IFN-a or IFN-b in these animals. Strong induction of IFN-ls by LIC-pIC occurred only in human hepatocytes, and not in mouse hepatocytes nor in human cell lines derived from other (non-hepatic) tissues. LIC-pIC-induced IFN-l production was mediated by the immune sensor adaptor molecules mitochondrial antiviral signaling protein (MAVS) and Toll/IL-1R domain-containing adaptor molecule-1 (TICAM-1), suggesting dual recognition of LIC-pIC by both sensor adaptor pathways. Conclusions: These findings demonstrate that the expression and function of various IFNs differ depending on the animal species and tissues under investigation. Chimeric mice harboring humanized livers demonstrate that IFN-ls play an important role in the defense against human hepatic virus infection.