scholarly journals Cytokine-Sensitive Replication of Hepatitis B Virus in Immortalized Mouse Hepatocyte Cultures

2002 ◽  
Vol 76 (11) ◽  
pp. 5646-5653 ◽  
Author(s):  
Valérie Pasquetto ◽  
Stefan F. Wieland ◽  
Susan L. Uprichard ◽  
Marco Tripodi ◽  
Francis V. Chisari
Keyword(s):  
Gene Expression ◽  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Intracellular Signaling ◽  
Growth Factor Receptor ◽  
Necrosis Factor Alpha ◽  
Tnf Α ◽  
B Virus ◽  
Ifn Γ

ABSTRACT We have previously shown that alpha/beta interferon (IFN-α/β) and gamma interferon (IFN-γ) 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-γ and tumor necrosis factor alpha (TNF-α) in the liver can inhibit HBV gene expression by destabilizing preformed viral mRNA. In order to further study the antiviral activity of IFN-α/β, IFN-γ, and TNF-α 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-α/β or IFN-γ, which eliminate pregenomic RNA-containing capsids from the cells as they do in the liver. Furthermore, we demonstrate that IFN-γ, but not IFN-α/β, is capable of inhibiting HBV gene expression in this system, especially when it acts synergistically with TNF-α. These cells should facilitate the analysis of the intracellular signaling pathways and effector mechanisms responsible for these antiviral effects.

scholarly journals Role of NF-κB and Myc Proteins in Apoptosis Induced by Hepatitis B Virus HBx Protein

2001 ◽  
Vol 75 (1) ◽  
pp. 215-225 ◽  
Author(s):  
Fei Su ◽  
Christian N. Theodosis ◽  
Robert J. Schneider
Keyword(s):  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Apoptotic Cell ◽  
Regulatory Protein ◽  
Family Members ◽  
Necrosis Factor Alpha ◽  
Tnf Α ◽  
B Virus ◽  
High Level

ABSTRACT Chronic infection with hepatitis B virus (HBV) promotes a high level of liver disease and cancer in humans. The HBV HBx gene encodes a small regulatory protein that is essential for viral replication and is suspected to play a role in viral pathogenesis. HBx stimulates cytoplasmic signal transduction pathways, moderately stimulates a number of transcription factors, including several nuclear factors, and in certain settings sensitizes cells to apoptosis by proapoptotic stimuli, including tumor necrosis factor alpha (TNF-α) and etopocide. Paradoxically, HBx activates members of the NF-κB transcription factor family, some of which are antiapoptotic in function. HBx induces expression of Myc protein family members in certain settings, and Myc can sensitize cells to killing by TNF-α. We therefore examined the roles of NF-κB, c-Myc, and TNF-α in apoptotic killing of cells by HBx. RelA/NF-κB is shown to be induced by HBx and to suppress HBx-mediated apoptosis. HBx also induces c-Rel/NF-κB, which can promote apoptotic cell death in some contexts or block it in others. Induction of c-Rel by HBx was found to inhibit its ability to directly mediate apoptotic killing of cells. Thus, HBx induction of NF-κB family members masks its ability to directly mediate apoptosis, whereas ablation of NF-κB reveals it. Investigation of the role of Myc protein demonstrates that overexpression of Myc is essential for acute sensitization of cells to killing by HBx plus TNF-α. This study therefore defines a specific set of parameters which must be met for HBx to possibly contribute to HBV pathogenesis.

scholarly journals Hepatitis B Virus RNA-Binding Proteins Associated with Cytokine-Induced Clearance of Viral RNA from the Liver of Transgenic Mice

1999 ◽  
Vol 73 (1) ◽  
pp. 474-481 ◽  
Author(s):  
Tilman Heise ◽  
Luca G. Guidotti ◽  
Victoria J. Cavanaugh ◽  
Francis V. Chisari
Keyword(s):  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Transgenic Mice ◽  
Rna Binding ◽  
Rna Binding Proteins ◽  
Viral Rna ◽  
Cytokine Activation ◽  
Tnf Α ◽  
B Virus ◽  
Ifn Γ

ABSTRACT Hepatitis B virus (HBV) gene expression is downregulated in the liver of HBV transgenic mice by a posttranscriptional mechanism that is triggered by the local production of gamma interferon (IFN-γ) and tumor necrosis factor alpha (TNF-α) during intrahepatic inflammation (hepatitis). The molecular basis for this antiviral effect is unknown. In this study, we identified three HBV RNA-binding liver nuclear proteins (p45, p39, and p26) the relative abundance of which correlates with the abundance of HBV RNA in response to the induction of IFN-γ and TNF-α. All three proteins bind to a 91-bp element located at the 5′ end of a previously defined posttranscriptional regulatory element that is thought to mediate the nuclear export of HBV RNA. The presence of p45 correlates directly with the presence of HBV RNA, being detectable under baseline conditions when the viral RNA is abundant and undetectable when the viral RNA disappears in response to IFN-γ and TNF-α. In contrast, p26 is inversely related to HBV RNA, being detectable only when the viral RNA disappears following cytokine activation. Finally, p39 is constitutively expressed, and its abundance and mobility appear to be slightly increased by cytokine activation. These results suggest a model in which hepatocellular HBV RNA content might be controlled by the stabilizing and/or destabilizing influences of these RNA-binding proteins whose activity is regulated by cytokine-induced signaling pathways.

scholarly journals Author Correction: Hypoxic gene expression in chronic hepatitis B virus infected patients is not observed in state-of-the-art in vitro and mouse infection models

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Peter Jianrui Liu ◽  
James M. Harris ◽  
Emanuele Marchi ◽  
Valentina D’Arienzo ◽  
Thomas Michler ◽  
...  
Keyword(s):  
Gene Expression ◽  
Chronic Hepatitis ◽  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Chronic Hepatitis B ◽  
State Of The Art ◽  
Infection Models ◽  
Chronic Hepatitis B Virus ◽  
B Virus

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

Συμβολή στη μελέτη των μεταλλαγών της πρωτεΐνης του πυρήνα του ιού της ηπατίτιδας Β και της αντιγονικής εκφράσεως σε ασθενείς με χρονία HBV λοίμωξη

2013 ◽  
Author(s):  
Ειρήνη Ράπτη
Keyword(s):  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Intracellular Cytokine Staining ◽  
Intracellular Cytokine ◽  
B Virus ◽  
Ifn Γ

Κυτταρικές ανοσιακές αντιδράσεις στην HBeAg αρνητική χρονία ηπατίτιδα ΒΣκοπός: Η ανοσοπαθογένεια της HBeAg αρνητικής χρονίας ηπατίτιδας Β (ΧΗΒ) δεν έχει μελετηθεί επαρκώς. Σκοπός λοιπόν της παρούσας μελέτης ήταν η αποτύπωση των κυτταρικών ανοσιακών αντιδράσεων στη μορφή αυτή της ΧΗΒ.Ασθενείς και μέθοδοι: Μελετήσαμε τις κυτταρικές ανοσιακές αντιδράσεις των περιφερικών λεμφοκυττάρων χρησιμοποιώντας λεμφοκυτταρικές καλλιέργειες, μεθόδους ενδοκυττάριας χρώσης κυττοκινών (intracellular cytokine staining (ICS) καθώς και μέτρησης της παραγωγής ιντερφερόνης-γ με τη μέθοδο ELISPOT μετά από μη ειδική και ειδική διέγερση με ολικές πρωτεΐνες του ιού της ηπατίτιδας Β (Hepatitis B Virus, HBV) αλλά και συνθετικά πεπτίδια.Τριάντα ασθενείς με HBeAg αρνητική ΧΗΒ, έντεκα ασυμπτωματικοί φορείς, εννέα ασθενείς με οξεία ηπατίτιδα Β και εικοσιδύο υγιείς μάρτυρες εισήχθησαν στη μελέτη.Αποτελέσματα: Οι ασθενείς με HBeAg (-) ΧΗΒ παρουσίασαν αυξημένο ποσοστό διεγερμένων περιφερικών CD8+ Τ-λεμφοκυττάρων και παράλληλα αυξημένο πολλαπλασιασμό των περιφερικών CD4 λεμφοκυττάρων μετά από in vitro διέγερση με αλληλοεπικαλυπτόμενα πεπτίδια του πυρηνοκαψιδίου του HBV καθώς και ένα πεπτίδιο του φακέλλου του HBV (HBs 182-191 aa), αντίδραση ομοιάζουσα σε ένταση με αυτή των ασθενών με οξεία ηπατίτιδα Β. Με τη χρήση ενδοκυττάριας χρώσης κυττοκινών (ICS), διαπιστώσαμε στην HBeAg (-) ΧΗΒ έναν διεγερμένο πληθυσμό Τ-λεμφοκυττάρων με παραγωγή IFN-γ (CD4+ και CD8+), μετά από μη ειδική διέγερση συγκριτικά με όλες τις άλλες ομάδες ασθενών και υγιών μαρτύρων. Επιπλέον οι ασθενείς με HBeAg(-) ΧΗΒ καθώς και αυτοί με οξεία ηπατίτιδα Β παρουσίασαν έναν εξίσου αυξημένο αριθμό ειδικών για το πυρηνοκαψίδιο του ιού (HBcore) T λεμφοκυττάρων, όπως μετρήθηκε με τις μεθόδους υπολογισμού της IFN-γ. Αντίθετα, ενώ οι ασυμπτωματικοί HBsAg φορείς είχαν ελάχιστες αντιδράσεις πολλαπλασιασμού των CD4+ λεμφοκυττάρων συνολικά όμως διατηρούσαν αυξημένο αριθμό διεγερμένων Τ-λεμφοκυττάρων ειδικών για την πρωτείνη του φακέλλου (όπως μετρήθηκε με ICS). Συμπεράσματα: Η μελέτη αυτή κατέδειξε ότι οι συνολικές CD4+ T λεμφοκυτταρικές αντιδράσεις των ασθενών με HBeAg (-) ΧΗΒ είναι συγκρίσιμες με αυτές των ασθενών με οξεία ηπατίτιδα Β, ενώ στους ασυμπτωματικούς HBsAg φορείς παρά τις περιορισμένες CD4+αντιδράσεις, η ικανότητα διέγερσης των περιφερικών Τ-λεμφοκυττάρων διατηρείται.

scholarly journals Tumor Necrosis Factor Alpha Inhibition of Hepatitis B Virus Replication Involves Disruption of Capsid Integrity through Activation of NF-κB

2003 ◽  
Vol 77 (7) ◽  
pp. 4033-4042 ◽  
Author(s):  
Michael Biermer ◽  
Robyn Puro ◽  
Robert J. Schneider
Keyword(s):  
Hepatitis B Virus ◽  
Tumor Necrosis Factor ◽  
Hepatitis B ◽  
Virus Replication ◽  
Tumor Necrosis ◽  
Necrosis Factor Alpha ◽  
Tnf Α ◽  
B Virus ◽  
Factor Alpha ◽  
Necrosis Factor

ABSTRACT Chronic infection by hepatitis B virus results from an inability to clear the virus, which is associated with liver disease and liver cancer. Tumor necrosis factor alpha (TNF-α) is associated with noncytopathic clearance of hepatitis B virus in animal models. Here we demonstrate that the nuclear factor κB (NF-κB) signaling pathway is a central mediator of inhibition of hepatitis B virus by TNF-α and we describe the molecular mechanism. TNF-α is shown to suppress hepatitis B virus DNA replication without cell killing by disrupting the formation or stability of cytoplasmic viral capsids through a pathway requiring the NF-κB-activating inhibitor of κB kinase complex IKK-α/β and active transcription factor NF-κB. Hepatitis B virus replication could also be inhibited and viral capsid formation could be disrupted in the absence of TNF-α solely by overexpression of IKK-α/β or strong activation of NF-κB. In contrast, inhibition of NF-κB signaling stimulated viral replication, demonstrating that HBV replication is both positively and negatively regulated by the level of activity of the NF-κB pathway. Studies are presented that exclude the possibility that HBV inhibition by NF-κB is carried out by secondary production of gamma interferon or alpha/beta interferon. These results identify a novel mechanism for noncytopathic suppression of hepatitis B virus replication that is mediated by the NF-κB signaling pathway and activated by TNF-α.

scholarly journals Inhibition of Hepatitis B Virus Replication during Adenovirus and Cytomegalovirus Infections in Transgenic Mice

1998 ◽  
Vol 72 (4) ◽  
pp. 2630-2637 ◽  
Author(s):  
Victoria J. Cavanaugh ◽  
Luca G. Guidotti ◽  
Francis V. Chisari
Keyword(s):  
Gene Expression ◽  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Transgenic Mice ◽  
Lymphocytic Choriomeningitis ◽  
Murine Cytomegalovirus ◽  
Necrosis Factor Alpha ◽  
Hbv Replication ◽  
B Virus ◽  
Cytomegalovirus Infections

ABSTRACT We have previously demonstrated that hepatitis B virus (HBV) replication and gene expression are abolished in the livers of HBV transgenic mice by cytotoxic T lymphocytes (CTLs) and during lymphocytic choriomeningitis virus (LCMV) infection, stimuli that trigger the production of alpha/beta interferon, gamma interferon, and tumor necrosis factor alpha in the liver. We now report that hepatic HBV replication and gene expression are inhibited by the local induction of these cytokines during adenovirus- and murine cytomegalovirus (MCMV)-induced hepatitis. Further, we show that MCMV also blocks HBV replication and gene expression in the proximal convoluted tubules of the kidney by causing interstitial nephritis and inducing the same cytokines in the renal parenchyma. These results suggest that inflammatory cytokines probably contribute to viral clearance during acute viral hepatitis in humans, and they imply that induction of these cytokines in the liver and other infected tissues of chronically infected patients might have therapeutic value.

scholarly journals Lack of Tumor Necrosis Factor Alpha Induces Impaired Proliferation of Hepatitis B Virus-Specific Cytotoxic T Lymphocytes

2003 ◽  
Vol 77 (4) ◽  
pp. 2469-2476 ◽  
Author(s):  
Senji Kasahara ◽  
Kazuki Ando ◽  
Kuniaki Saito ◽  
Kenji Sekikawa ◽  
Hiroyasu Ito ◽  
...  
Keyword(s):  
Hepatitis B Virus ◽  
Tumor Necrosis Factor ◽  
Cytotoxic T Lymphocytes ◽  
Tumor Necrosis ◽  
Necrosis Factor Alpha ◽  
Tnf Α ◽  
B Virus ◽  
Factor Alpha ◽  
Necrosis Factor

ABSTRACT Recent studies have shown that tumor necrosis factor alpha (TNF-α) plays critical roles in not only viral clearance but also lymphoid tissue development and stem cell differentiation. In this study, we attempted to induce hepatitis B virus (HBV)-specific cytotoxic T lymphocytes (CTLs) by immunization of TNF-α knockout (TNF-α−/−) mice with HBsAg-encoding plasmid DNA. An immunization with the HBV plasmid failed to induce CTL responses in TNF-α−/− mice, although CTLs were readily induced in wild-type mice by the same protocol. Weak CTL responses were produced in TNF-α−/− mice after two sessions of immunization with the HBV plasmid; however, TNF-α was required to maintain the responses of these CTL lines to in vitro stimulation and, even then, the responses were lost after 3 weeks. Interestingly, a limiting dilution of a CTL line showed that HBV-specific CTL clones with high specific cytotoxicity were present in TNF-α−/− mice, but these clones again failed to proliferate for more than 3 weeks. Furthermore, since exogenously added TNF-α enhanced the proliferation of a TNF-α−/− clone but suppressed that of a TNF-α+/+ clone in vitro, TNF-α also has a direct effect on the proliferation of CTLs. In conclusion, TNF-α is essential rather than important for the proliferation of HBV-specific CTLs both in vivo and in vitro and this effect is not only due to the activation of dendritic cells but is also induced by the direct effect on CTLs.

scholarly journals Indispensable Role for TNF-α and IFN-γ at the Effector Phase of Liver Injury Mediated by Th1 Cells Specific to Hepatitis B Virus Surface Antigen

2000 ◽  
Vol 165 (2) ◽  
pp. 956-961 ◽  
Author(s):  
Akio Ohta ◽  
Masashi Sekimoto ◽  
Marimo Sato ◽  
Toshiaki Koda ◽  
Shin-ichiro Nishimura ◽  
...  
Keyword(s):  
Hepatitis B Virus ◽  
Liver Injury ◽  
Hepatitis B ◽  
Surface Antigen ◽  
Th1 Cells ◽  
Virus Surface ◽  
Effector Phase ◽  
Tnf Α ◽  
B Virus ◽  
Ifn Γ
Sign in / Sign up

Export Citation Format

Share Document