Macromolecular synthesis in hepatitis A virus-infected cells

1986 ◽  
Vol 42 (1) ◽  
pp. 91-91
Author(s):  
J. de Chastonay ◽  
G. Siegl
Keyword(s):  
Hepatitis A ◽  
Hepatitis A Virus ◽  
Macromolecular Synthesis ◽  
Infected Cells

scholarly journals Heme Oxygenase-1 Exerts Antiviral Activity against Hepatitis A Virus In Vitro

Pharmaceutics ◽  
2021 ◽  
Vol 13 (8) ◽  
pp. 1229
Author(s):  
Dong-Hwi Kim ◽  
Hee-Seop Ahn ◽  
Hyeon-Jeong Go ◽  
Da-Yoon Kim ◽  
Jae-Hyeong Kim ◽  
...  
Keyword(s):  
Antiviral Activity ◽  
Protein Expression ◽  
Heme Oxygenase ◽  
Hepatitis A ◽  
Hepatitis A Virus ◽  
Health Concern ◽  
Heme Oxygenase 1 ◽  
Infected Cells ◽  
In Cells

Hepatitis A virus (HAV), the causative pathogen of hepatitis A, induces severe acute liver injuries in humans and is a serious public health concern worldwide. However, appropriate therapeutics have not yet been developed. The enzyme heme oxygenase-1 (HO-1) exerts antiviral activities in cells infected with several viruses including hepatitis B and C viruses. In this study, we demonstrated for the first time the suppression of virus replication by HO-1 in cells infected with HAV. Hemin (HO-1 inducer) induced HO-1 mRNA and protein expression, as expected, and below 50 mM, dose-dependently reduced the viral RNA and proteins in the HAV-infected cells without cytotoxicity. Additionally, HO-1 protein overexpression using a protein expression vector suppressed HAV replication. Although ZnPP-9, an HO-1 inhibitor, did not affect HAV replication, it significantly inhibited hemin-induced antiviral activity in HAV-infected cells. Additionally, FeCl3, CORM-3, biliverdin, and the HO-1 inducers andrographolide and CoPP inhibited HAV replication in the HAV-infected cells; andrographolide and CoPP exhibited a dose-dependent effect. In conclusion, these results suggest that HO-1 effectively suppresses HAV infection in vitro, and its enzymatic products appear to exert antiviral activity. We expect that these results could contribute to the development of a new antiviral drug for HAV.

scholarly journals Detection of Hepatitis A Virus by Using a Combined Cell Culture-Molecular Beacon Assay

2008 ◽  
Vol 74 (7) ◽  
pp. 2239-2243 ◽  
Author(s):  
Hsiao-Yun Yeh ◽  
Yu-Chen Hwang ◽  
Marylynn V. Yates ◽  
Ashok Mulchandani ◽  
Wilfred Chen
Keyword(s):  
Cell Culture ◽  
Hepatitis A ◽  
Molecular Beacon ◽  
Hepatitis A Virus ◽  
Mammalian Cell Culture ◽  
Fluorescence Signal ◽  
Cytopathic Effects ◽  
Infected Cells ◽  
Fetal Rhesus Monkey Kidney ◽  
Fluorescent Cells

ABSTRACT Rapid and efficient methods for the detection and quantification of infectious viruses are required for public health risk assessment. Current methods to detect infectious viruses are based on mammalian cell culture and rely on the production of visible cytopathic effects (CPE). For hepatitis A virus (HAV), viral replication in cell culture has been reported to be nonlytic and relatively slow. It may take more than 1 week to reach the maximum production and subsequent visualization of CPE. A molecular beacon (MB), H1, specifically targeting a 20-bp 5′ noncoding region of HAV, was designed and synthesized. MB H1 was introduced into fixed and permeabilized fetal rhesus monkey kidney (FRhK-4) cells infected with HAV strain HM-175. Upon hybridizing with the viral mRNA, fluorescent cells were visualized easily under a fluorescence microscope. Discernible fluorescence was detected only in infected cells by using the specific MB H1. A nonspecific MB, which was not complementary to the viral RNA sequence, produced no visible fluorescence signal. This MB-based fluorescence assay enabled the direct counting of fluorescent cells and could achieve a detection limit of 1 PFU at 6 h postinfection, demonstrating a significant improvement in viral quantification over current infectivity assays.

scholarly journals CXCL10 is produced in hepatitis A virus-infected cells in an IRF3-dependent but IFN-independent manner

2017 ◽  
Vol 7 (1) ◽  
Author(s):  
Pil Soo Sung ◽  
Seon-Hui Hong ◽  
Jeewon Lee ◽  
Su-Hyung Park ◽  
Seung Kew Yoon ◽  
...  
Keyword(s):  
Hepatitis A ◽  
Hepatitis A Virus ◽  
Independent Manner ◽  
Infected Cells

Hepatitis A virus-encoded miRNAs attenuate the accumulation of viral genomic RNAs in infected cells

Virus Genes ◽  
2016 ◽  
Vol 52 (3) ◽  
pp. 317-324 ◽  
Author(s):  
Jiandong Shi ◽  
Jing Sun ◽  
Meini Wu ◽  
Ningzhu Hu ◽  
Yunzhang Hu
Keyword(s):  
Hepatitis A ◽  
Hepatitis A Virus ◽  
Genomic Rnas ◽  
Infected Cells

Perinuclear accumulation of hepatitis A virus proteins, RNA, and particles and ultrastructural alterations in infected cells

2001 ◽  
Vol 146 (12) ◽  
pp. 2291-2307 ◽  
Author(s):  
M. H. F. Klinger ◽  
R. Kämmerer ◽  
B. Hornei ◽  
V. Gauss-Müller
Keyword(s):  
Hepatitis A ◽  
Hepatitis A Virus ◽  
Ultrastructural Alterations ◽  
Infected Cells ◽  
Virus Proteins

scholarly journals Cellular entry and uncoating of naked and quasi-enveloped human hepatoviruses

eLife ◽  
2019 ◽  
Vol 8 ◽  
Author(s):  
Efraín E Rivera-Serrano ◽  
Olga González-López ◽  
Anshuman Das ◽  
Stanley M Lemon
Keyword(s):  
Hepatitis A ◽  
Hepatitis A Virus ◽  
Surface Proteins ◽  
Integrin Β1 ◽  
Enveloped Viruses ◽  
Environmental Transmission ◽  
Late Endosomes ◽  
Infected Cells ◽  
Cell Spread ◽  
Endocytic Pathways

Many ‘non-enveloped’ viruses, including hepatitis A virus (HAV), are released non-lytically from infected cells as infectious, quasi-enveloped virions cloaked in host membranes. Quasi-enveloped HAV (eHAV) mediates stealthy cell-to-cell spread within the liver, whereas stable naked virions shed in feces are optimized for environmental transmission. eHAV lacks virus-encoded surface proteins, and how it enters cells is unknown. We show both virion types enter by clathrin- and dynamin-dependent endocytosis, facilitated by integrin β1, and traffic through early and late endosomes. Uncoating of naked virions occurs in late endosomes, whereas eHAV undergoes ALIX-dependent trafficking to lysosomes where the quasi-envelope is enzymatically degraded and uncoating ensues coincident with breaching of endolysosomal membranes. Neither virion requires PLA2G16, a phospholipase essential for entry of other picornaviruses. Thus naked and quasi-enveloped virions enter via similar endocytic pathways, but uncoat in different compartments and release their genomes to the cytosol in a manner mechanistically distinct from other Picornaviridae.

scholarly journals Silencing of Hepatitis A Virus Infection by Small Interfering RNAs

2006 ◽  
Vol 80 (11) ◽  
pp. 5599-5610 ◽  
Author(s):  
Yuri Kusov ◽  
Tatsuo Kanda ◽  
Ann Palmenberg ◽  
Jean-Yves Sgro ◽  
Verena Gauss-Müller
Keyword(s):  
Hepatitis A ◽  
Hepatitis A Virus ◽  
Firefly Luciferase ◽  
Viral Escape ◽  
Small Interfering Rnas ◽  
Therapeutic Tool ◽  
Escape Mutants ◽  
Infected Cells ◽  
Multiple Sequences ◽  
Insight Into

ABSTRACT Infection by hepatitis A virus (HAV) can cause acute hepatitis and, rarely, fulminant liver failure, in particular in patients chronically infected with hepatitis C virus. Based on our previous observation that small interfering RNAs (siRNAs) can silence translation and replication of the firefly luciferase-encoding HAV replicon, we now exploited this technology to demonstrate the effect of siRNAs on viral infection in Huh-7 cells. Freshly and persistently infected cells were transfected with siRNAs targeting various sites in the HAV nonstructural genes. Compared to a single application, consecutive siRNA transfections targeting multiple sequences in the viral genome resulted in a more efficient and sustained silencing effect than a single transfection. In most instances, multiple applications of a single siRNA led to the emergence of viral escape mutants with mutated target sites that rendered these genomes resistant to RNA interference (RNAi). Efficient and sustained suppression of the viral infectivity was achieved after consecutive applications of an siRNA targeting a computer-predicted hairpin structure. This siRNA holds promise as a therapeutic tool for severe courses of HAV infection. In addition, the results provide new insight into the structural bases for sequence-specific RNAi.

scholarly journals Characterization of Replication-Competent Hepatitis A Virus Constructs Containing Insertions at the N Terminus of the Polyprotein

1998 ◽  
Vol 72 (1) ◽  
pp. 349-357 ◽  
Author(s):  
Yuan Zhang ◽  
Gerardo G. Kaplan
Keyword(s):  
Western Blot ◽  
Blot Analysis ◽  
Cleavage Site ◽  
Western Blot Analysis ◽  
Hepatitis A ◽  
Hepatitis A Virus ◽  
Flag Epitope ◽  
Initiation Of Translation ◽  
N Terminus ◽  
Infected Cells

ABSTRACT To determine whether hepatitis A virus (HAV) could tolerate the insertion of exogenous sequences, we constructed full-length HAV cDNAs containing in-frame insertions at the N terminus of the polyprotein and transfected the derived T7 RNA polymerase in vitro transcripts into FRhK-4 cells. Replication of HAVvec1, a construct containing an insertion of 60 nucleotides coding for a polylinker, a 2B/2C cleavage site for HAV protease 3Cpro, and two initiation codons that restored the sequence of the N terminus of the polyprotein, was detected 2 weeks after transfection by indirect immunofluorescence analysis using anti-HAV monoclonal antibodies. Western blot analysis of HAVvec1-infected cells using anti-VP2 and anti-VP4 antibodies failed to detect the expression of the inserted sequences. Insertion of a 24-mer oligonucleotide coding for a FLAG epitope into HAVvec1 resulted in its HAV-mediated expression which was retained upon deletion of a Gln residue from the inserted 2B/2C cleavage site. Western blot analysis using anti-FLAG and anti-VP2 antibodies showed that the FLAG epitope accumulated in infected cells fused to VP0. Replacement of the FLAG epitope with an epitope of the circumsporozoite protein (CSP) ofPlasmodium falciparum resulted in its stable HAV-mediated expression for at least six serial passages in FRhK-4 cells. Sedimentation analysis in sucrose density gradients showed that the CSP epitope accumulated in infected cells fused to VP0, forming 80S empty capsids which also contained native VP0. Our data suggest that the HAV internal ribosome entry site can efficiently direct dual initiation of translation of the polyprotein from AUG codons separated by 66 to 78 nucleotides and show that HAV can tolerate insertions at the N terminus of the polyprotein.

Sign in / Sign up

Export Citation Format

Share Document