scholarly journals Release of Genotype 1 Hepatitis E Virus from Cultured Hepatoma and Polarized Intestinal Cells Depends on Open Reading Frame 3 Protein and Requires an Intact PXXP Motif

2010 ◽  
Vol 84 (18) ◽  
pp. 9059-9069 ◽  
Author(s):  
Suzanne U. Emerson ◽  
Hanh T. Nguyen ◽  
Udana Torian ◽  
Danielle Burke ◽  
Ronald Engle ◽  
...  

ABSTRACT Hepatitis E virus genotype 1 strain Sar55 replicated in subcloned Caco-2 intestinal cells and Huh7 hepatoma cells that had been transfected with in vitro transcribed viral genomes, and hepatitis E virions were released into the culture medium of both cell lines. Virus egress from cells depended on open reading frame 3 (ORF3) protein, and a proline-rich sequence in ORF3 was important for egress from cultured cells and for infection of macaques. Both intracellular ORF3 protein accumulation and virus release occurred at the apical membrane of polarized Caco-2 cells. ORF3 protein and lipids were intimately associated with virus particles produced in either cell line; ORF2 epitopes were masked in these particles and could not be immunoprecipitated with anti-ORF2.

2009 ◽  
Vol 1 (2) ◽  
pp. 77-84 ◽  
Author(s):  
Nereida Jiménez de Oya ◽  
Inmaculada Galindo ◽  
Estela Escribano-Romero ◽  
Ana-Belén Blázquez ◽  
Julio Alonso-Padilla ◽  
...  

2005 ◽  
Vol 79 (11) ◽  
pp. 6680-6689 ◽  
Author(s):  
Judith Graff ◽  
Hanh Nguyen ◽  
Claro Yu ◽  
William R. Elkins ◽  
Marisa St. Claire ◽  
...  

ABSTRACT An infectious cDNA clone of hepatitis E virus was mutated in order to prevent synthesis of either open reading frame 2 (ORF2) protein or ORF3 protein. HuH-7 cells transfected with an ORF2-null mutant produced ORF3, and those transfected with an ORF3-null mutant produced ORF2. Silent mutations introduced into a highly conserved nucleotide sequence in the ORF3 coding region eliminated the synthesis of both ORF2 and ORF3 proteins, suggesting that it comprised a cis-reactive element. A mutant that was not able to produce ORF3 protein did not produce a detectable infection in rhesus macaques. However, a mutant that encoded an ORF3 protein lacking a phosphorylation site reported to be critical for function was able to replicate its genome in cell culture and to induce viremia and seroconversion in rhesus monkeys, suggesting that phosphorylation of ORF3 protein was not necessary for genome replication or for production of infectious virions.


Author(s):  
M. I. Mikhailov ◽  
E. Yu. Malinnikova ◽  
K. K. Kyuregyan ◽  
O. V. Isaeva

Aim. Description of the first documented case of imported hepatitis E, associated with genotype 4 of HEV and introduced from southern France. Materials and methods. Clinical, epidemiologic and laboratory analysis of the imported case of disease of hepatitis E was carried out. Phylogenetic analysis of nucleotide sequences of HEV isolate, taken from the patient, was carried out. Results. Epidemiologic analysis allowed to assume imported character of the detected case of HEV-infection. Comparative analysis of nucleotide sequences of regions of the open reading frame 2 (300 nt) and open reading frame 1 (721 nt) of HEV genome, isolated from the patient, showed identity of this isolate with variants of genotype 4 HEV, isolated in France in 2009 - 2011 from patients with autochthonous hepatitis E. Conclusion. The results obtained confirm the case of import into Russia of genotype 4 HEV from south-eastern France (Corsica), where spread of this virus genotype is observed in recent years.


2019 ◽  
Vol 8 (1) ◽  
Author(s):  
Olusola Anuoluwapo Akanbi ◽  
Dominik Harms ◽  
Bo Wang ◽  
Oluyinka Oladele Opaleye ◽  
Olufisayo Adesina ◽  
...  

Hepatitis E virus genotype 1 (HEV-1) is associated with large epidemics. Notably, HEV subtype 1e (HEV-1e) has caused HEV outbreaks in sub-Saharan Africa.


Virulence ◽  
2021 ◽  
Vol 12 (1) ◽  
pp. 1334-1344
Author(s):  
Mohamed A. El-Mokhtar ◽  
Haidi Karam-Allah Ramadan ◽  
Muhamad R. Abdel Hameed ◽  
Ayat M. Kamel ◽  
Sahar A. Mandour ◽  
...  

Viruses ◽  
2021 ◽  
Vol 13 (7) ◽  
pp. 1385
Author(s):  
Giulia Pezzoni ◽  
Lidia Stercoli ◽  
Eleonora Pegoiani ◽  
Emiliana Brocchi

To evaluate the antigenic properties of Hepatitis E Virus (HEV) Open Reading Frame 2 and 3 (ORF2 and ORF3) codified proteins, we expressed different portions of ORF2 and the entire ORF3 in E. coli, a truncated ORF2, was also expressed in baculovirus. A panel of 37 monoclonal antibodies (MAbs) was raised against ORF2 (1–660 amino acids) and MAbs were mapped and characterized using the ORF2 expressed portions. Selected HEV positive and negative swine sera were used to evaluate ORF2 and ORF3 antigens’ immunogenicity. The MAbs were clustered in six groups identifying six antigenic regions along the ORF2. Only MAbs binding to the sixth ORF2 antigenic region (394–608 aa) were found to compete with HEV positive sera and efficiently catch the recombinant antigen expressed in baculovirus. The ORF2 portion from 394–608 aa demonstrated to include most immunogenic epitopes with 85% of HEV positive swine sera reacting against the region from 461–544 aa. Only 5% of the selected HEV sera reacted against the ORF3 antigen.


2013 ◽  
Vol 86 (3) ◽  
pp. 487-495 ◽  
Author(s):  
Yan Zhou ◽  
Yansheng Geng ◽  
Jun Yang ◽  
Chenyan Zhao ◽  
Tim J. Harrison ◽  
...  

PLoS ONE ◽  
2020 ◽  
Vol 15 (9) ◽  
pp. e0237618
Author(s):  
Youkyung H. Choi ◽  
Xiugen Zhang ◽  
Ganesh Srinivasamoorthy ◽  
Michael A. Purdy

2019 ◽  
Vol 220 (5) ◽  
pp. 811-819 ◽  
Author(s):  
Ibrahim M Sayed ◽  
Lieven Verhoye ◽  
Claire Montpellier ◽  
Florence Abravanel ◽  
Jacques Izopet ◽  
...  

Abstract Background Hepatitis E virus infection (HEV) is an emerging problem in developed countries. Diagnosis of HEV infection is based on the detection of HEV-specific antibodies, viral RNA, and/or antigen (Ag). Humanized mice were previously reported as a model for the study of HEV infection, but published data were focused on the quantification of viral RNA. However, the kinetics of HEV Ag expression during infection remains poorly understood. Methods Plasma specimens and suspensions of fecal specimens from HEV-infected and ribavirin-treated humanized mice were analyzed using HEV antigen–specific enzyme-linked immunosorbent assay, reverse transcription–quantitative polymerase chain reaction analysis, density gradient analysis, and Western blotting. Result Open reading frame 2 (ORF2) Ag was detected in both plasma and stool from HEV-infected mice, and levels increased over time. Contrary to HEV RNA, ORF2 Ag levels were higher in mouse plasma than in stool. Interestingly, ORF2 was detected in plasma from mice that tested negative for HEV RNA in plasma but positive for HEV RNA in stool and was detected after viral clearance in mice that were treated with ribavirin. Plasma density gradient analysis revealed the presence of the noninfectious glycosylated form of ORF2. Conclusion ORF2 Ag can be used as a marker of active HEV infection and for assessment of the effect of antiviral therapy, especially when fecal samples are not available or molecular diagnostic tests are not accessible.


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