Isolation and identification of hepatitis E virus in Xinjiang, China

Huang, Rutong; Li, D. R.; Wei, Jinbei; Huang, Xing‐Yao; Yuan, Xitong; Xue, Tian

doi:10.1099/0022-1317-73-5-1143

Cited by 64 publications

(35 citation statements)

References 11 publications

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“…This region has also been shown to carry an immunodominant epitope, antibodies to which are universally present in infected humans and animals NOTES 12085 (24). Moreover, this region is highly conserved in all HEV strains sequenced to date (2,4,24,32,35,41) except the Mexican strain (14). However, our experiments with the ORF3 from the Mexican strain of HEV indicated that despite the variability in the sequence, the Mexican strain ORF3 still interacts with bikunin in the two-hybrid system.…”

Section: Hismentioning

confidence: 54%

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The 41-Amino-Acid C-Terminal Region of the Hepatitis E Virus ORF3 Protein Interacts with Bikunin, a Kunitz-Type Serine Protease Inhibitor

Tyagi

Surjit

Lal

2005

J Virol

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Hepatitis E virus (HEV), a human plus-stranded RNA virus, contains three open reading frames (ORF).Of these, ORF1 encodes the viral nonstructural polyprotein, ORF2 encodes the major capsid protein, and ORF3 codes for a phosphoprotein of undefined function. Recently, using the yeast two-hybrid system to screen a human cDNA liver library, we have isolated and characterized AMBP (␣ 1 -microglobulin/bikunin precursor), which specifically interacts with the ORF3 protein of HEV. The ORF3 protein expedites the processing and secretion of ␣ 1 -microglobulin. When checked individually for interaction, the second processed protein from AMBP, bikunin, strongly interacted with the full-length ORF3 protein. This protein-protein interaction has been validated by immunoprecipitation in both COS-1 and Huh7 cells and by His 6 pull-down assays. In dual-labeling immunofluorescent staining, followed by fluorescence microscopy of transfected human liver cells, ORF3 colocalized with endogenously expressed bikunin. Finally, a 41-amino-acid C-terminal region of ORF3 has been found to be responsible for interacting with bikunin. The importance of this virus-host protein-protein interaction, with reference to the viral life cycle, has been discussed.Hepatitis E is an acute disease endemic in many countries throughout developing parts of the world, in particular on the continents of Africa and Asia, where it causes epidemics and sporadic infections. The causative agent, hepatitis E virus (HEV), is transmitted via the fecal-oral route, predominantly through contaminated water (7,10,25,26). HEV is a plusstranded RNA virus with a 7.2-kb genome containing three open reading frames (ORF), ORF1, ORF2, and ORF3, encoding three different proteins (20,32,35). ORF1 (5,079 bp) is at the 5Ј end of the genome and is predicted to code for the putative nonstructural proteins with sequences homologous to those encoding viral methyltransferases, proteases, helicases, and RNA-dependent RNA polymerases (1,20,27,35). In the absence of a reliable in vitro culture system for HEV, fundamental studies on its replication and expression strategy have not been undertaken. ORF2 and ORF3 have been expressed in Escherichia coli, animal cells, baculovirus, and yeast and in vitro in a coupled transcription-translation system (12,15,17,22,23). ORF2, the major capsid protein for HEV, is synthesized as a precursor and is processed through signal sequence cleavage into the mature protein, which is capable of selfassociation (18, 36). ORF3 encodes a small 13.5-kDa phosphoprotein that is expressed intracellularly, associates with the cytoskeleton, and shows no major processing (3, 42). The ORF3 protein dimerizes using a 43-amino-acid region, interacts with SH3 domains, and activates mitogen-activated protein kinase (21, 37). The phosphorylated form of the ORF3 protein has also been shown to interact with the nonglycosylated form of the major capsid protein, ORF2 (38). These properties of ORF3 clearly indicate that this protein may have multiple roles in HEV pathogenesis. Rece...

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Section: Hismentioning

confidence: 54%

“…The C-terminal region of ORF3 is highly conserved in all HEV strains, except in the Mexican strain (14). The ORF3 from the Mexican strain of HEV (BD-Mex ORF3) was also tested for interaction with AD-bikunin and showed growth on the SDTrp…”

Section: Hismentioning

confidence: 99%

The 41-Amino-Acid C-Terminal Region of the Hepatitis E Virus ORF3 Protein Interacts with Bikunin, a Kunitz-Type Serine Protease Inhibitor

Tyagi

Surjit

Lal

2005

J Virol

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“…The outbreak lasted for over 2 years and was associated with continuous drinking-water contamination. 20 In recent years, swine HEV isolates (genotype 4) were also identified in this region. 21,22 However, the significant genetic diversity between the swine and human HEV isolates suggested that the swine HEV was unlikely to be the major source of the hepatitis E outbreak in Xinjiang.…”

Section: Hev As the Sole Member Of The Hepevirus Genus Inmentioning

confidence: 99%

Hepatitis E Virus Infection among Animals and Humans in Xinjiang, China: Possibility of Swine to Human Transmission of Sporadic Hepatitis E in an Endemic Area

Li²,

Zhu³

et al. 2010

The American Society of Tropical Medicine and Hygiene

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Abstract. Hepatitis E is a worldwide public health problem, especially in areas with poor sanitation. This study examines the potential hepatitis E virus (HEV) animal reservoirs and the current status of HEV infection among animals and humans in an endemic area of Xinjiang, China. One thousand five hundred twenty-one serum samples from 12 different animal species and 296 sera from humans were detected for anti-HEV with an in-house enzyme immunoassay, and partial HEV RNA was amplified with a reverse transcription-nested polymerase chain reaction (RT-nPCR). All these distinct animal species, except jerboa and hoptoad, were positive for anti-HEV. However, HEV RNA was only amplified from pigs and a sporadic hepatitis E case in humans. The human HEV strain (CHN-XJ-HE29) shared 100% nucleotide identity with the swine HEV strain (CHN-XJ-SW50), both of which were collected from the same district; this indicates the possibility of HEV transmission from swine to humans in an endemic area.

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“…Propagation of HEV in vitro has been attempted in various cell lines (Divizia et al, 1999;Huang et al, 1992;Kazachkov et al, 1992;Meng et al, 1997;Wei et al, 2000), but an efficient cell culture system has not been developed. Several research groups have established infectious cDNA clones of HEV, which were observed to replicate in non-human primates or pigs (Emerson et al, 2001(Emerson et al, , 2004bGraff et al, 2005;Huang et al, 2005Huang et al, , 2007Panda et al, 2000).…”

mentioning

confidence: 99%

Construction of an infectious cDNA clone of hepatitis E virus strain JE03-1760F that can propagate efficiently in cultured cells

Yamada

Takahashi

Hoshino

et al. 2009

Journal of General Virology

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A full-length infectious cDNA clone (pJE03-1760F/wt) of a genotype 3 hepatitis E virus (HEV) (strain JE03-1760F) obtained from a faecal specimen was constructed in this study. Upon transfection of the capped in vitro transcripts of pJE03-1760F/wt into PLC/PRF/5 cells, the viral RNA levels in the culture supernatant started to increase on day 6 post-transfection (p.t.) and reached 107 copies ml−1 on day 28 p.t. Detection of increasing numbers of cells with ORF2 protein expression by immunofluorescence assay at 5, 7, 11 and 15 days p.t. indicated the spread of HEV infection in cell culture. When the cDNA-derived virus in culture supernatant was inoculated into PLC/PRF/5 or A549 cells, it grew as efficiently as the faeces-derived virus in both cells, reaching 106 copies ml−1 at 30 days post-inoculation. Our reverse genetics system for HEV that is usable in a robust cell-culture system will be useful for elucidation of the mechanism of HEV replication and functional roles of HEV proteins.

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Isolation and identification of hepatitis E virus in Xinjiang, China

Cited by 64 publications

References 11 publications

The 41-Amino-Acid C-Terminal Region of the Hepatitis E Virus ORF3 Protein Interacts with Bikunin, a Kunitz-Type Serine Protease Inhibitor

The 41-Amino-Acid C-Terminal Region of the Hepatitis E Virus ORF3 Protein Interacts with Bikunin, a Kunitz-Type Serine Protease Inhibitor

Hepatitis E Virus Infection among Animals and Humans in Xinjiang, China: Possibility of Swine to Human Transmission of Sporadic Hepatitis E in an Endemic Area

Construction of an infectious cDNA clone of hepatitis E virus strain JE03-1760F that can propagate efficiently in cultured cells

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