Genotype 5 Hepatitis E Virus Produced by a Reverse Genetics System Has the Potential for Zoonotic Infection

Bactrian camel hepatitis E virus (HEV) is a novel HEV belonging to genotype 8 (HEV-8) in the Orthohepevirus A species of the genus Hepevirus in the family Hepeviridae. HEV-8 cross-transmits to cynomolgus monkeys and has a potential risk for zoonotic infection. Until now, neither a cell-culture system to grow the virus nor a reverse genetics system to generate the virus has been developed. To generate replication-competent HEV-8 and to establish a cell-culture system, we synthesized capped genomic HEV-8 RNAs by in vitro transcription and used them to transfect into PLC/PRF/5 cells. A HEV-8 strain, HEV-8M2, was recovered from the capped HEV-8 RNA–transfected cell-culture supernatants and subsequently passaged in the cells, demonstrating that PLC/PRF/5 cells were capable of supporting the replication of the HEV-8, and that a cell-culture system for HEV-8 was successfully established. In addition to PLC/PRF/5 cells, A549 and Caco-2 cells appeared to be competent for the replication, but HepG2 C3/A, Vero, Hela S3, HEp-2C, 293T and GL37 cells were incompetent. The HEV-8M2 strain was capable of infecting cynomolgus monkeys by an intravenous inoculation, indicating that HEV-8 was infectious and again carried a risk for zoonotic infection. In contrast, HEV-8 did not infect nude rats and BALB/c nude mice, suggesting that the reservoir of HEV-8 was limited. In addition, the replication of the HEV-8M2 strain was efficiently abrogated by ribavirin but not by favipiravir, suggesting that ribavirin is a drug candidate for therapeutic treatment of HEV-8-induced hepatitis. The infectious HEV-8 produced by a reverse genetics system would be useful to elucidate the mechanisms of HEV replication and the pathogenesis of type E hepatitis.

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Successful infection of BALB/c mice by a swine hepatitis E virus clone constructed with reverse genetics

BMC Infectious Diseases ◽

10.1186/s12879-018-3544-4 ◽

2018 ◽

Vol 18 (1) ◽

Cited By ~ 6

Author(s):

Wenhai Yu ◽

Chenchen Yang ◽

Xianhui Hao ◽

Tianwu Ma ◽

Fen Huang

Keyword(s):

Hepatitis E Virus ◽

Reverse Genetics ◽

Hepatitis E ◽

Successful Infection ◽

Swine Hepatitis ◽

Virus Clone

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From barnyard to food table: The omnipresence of hepatitis E virus and risk for zoonotic infection and food safety

Virus Research ◽

10.1016/j.virusres.2011.01.016 ◽

2011 ◽

Vol 161 (1) ◽

pp. 23-30 ◽

Cited By ~ 207

Author(s):

Xiang-Jin Meng

Keyword(s):

Food Safety ◽

Hepatitis E Virus ◽

Hepatitis E ◽

Zoonotic Infection

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A Cross-Species Transmission of a Camel-Derived Genotype 8 Hepatitis E Virus to Rabbits

Pathogens ◽

10.3390/pathogens10111374 ◽

2021 ◽

Vol 10 (11) ◽

pp. 1374

Author(s):

Wenjing Zhang ◽

Yasushi Ami ◽

Yuriko Suzaki ◽

Michiyo Kataoka ◽

Naokazu Takeda ◽

...

Keyword(s):

Neutralizing Antibodies ◽

Hepatitis E Virus ◽

Hepatitis E ◽

Igg Antibodies ◽

Zoonotic Infection ◽

Significant Elevation ◽

Dromedary Camels ◽

Cynomolgus Monkeys ◽

Natural Reservoir ◽

Bactrian Camels

Novel genotypes of hepatitis E virus (HEV), i.e., HEV-5, HEV-7, and HEV-8, have been identified in wild boar, dromedary camels, and Bactrian camels, respectively, and they transmit to cynomolgus monkeys in a trans-species manner, raising the potential for zoonotic infection. Rabbits are the natural reservoir for rabbit HEV, but they are also susceptible to HEV-3 and HEV-4. It has been unknown whether rabbits are susceptible to HEV-5, HEV-7, and HEV-8. To investigate the infectivity of novel HEVs in rabbits and to assess whether rabbits are appropriate animal models for these HEVs, we inoculated Japanese white rabbits with HEV-5, HEV-7, and HEV-8, respectively. We observed that viral RNA was present in the fecal specimens of the HEV-8-inoculated rabbits and anti-HEV IgG antibodies were present in its sera, although anti-HEV IgM was undetectable and no significant elevation of ALT was observed. These results indicated that HEV-8 crossed species and infected the rabbits. No evidence for replication was observed in HEV-5 and HEV-7, suggesting that rabbits are not susceptible to these genotypes. The antibodies elicited in the HEV-8-infected rabbits did not protect them from the rabbit HEV challenge, suggesting that the antigenicity differs between HEV-8 and rabbit HEV. Antigenic analyses demonstrated that anti-HEV-8 antibodies reacted more strongly with homologous HEV-8 virus-like particles (VLPs) compared to heterologous rabbit HEV VLPs, but anti-rabbit HEV antibody had similar reactivity to the VLPs of rabbit HEV and HEV-8, suggesting that HEV-8 lacks some epitope(s) that exist in rabbit HEV and induced the neutralizing antibodies against rabbit HEV.

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Establishment of a Plasmid-Based Reverse Genetics System for the Cell Culture-Adapted Hepatitis E Virus Genotype 3c Strain 47832c

Pathogens ◽

10.3390/pathogens9030157 ◽

2020 ◽

Vol 9 (3) ◽

pp. 157 ◽

Cited By ~ 4

Author(s):

Johannes Scholz ◽

Christine Bächlein ◽

Ashish K. Gadicherla ◽

Alexander Falkenhagen ◽

Simon H. Tausch ◽

...

Keyword(s):

Cell Culture ◽

Hepatitis E Virus ◽

Reverse Genetics ◽

Single Point ◽

Hepatitis E ◽

Site Directed Mutagenesis ◽

Additional Restriction ◽

Progeny Virus ◽

Single Point Mutation ◽

Directed Mutagenesis

The hepatitis E virus (HEV) causes acute and chronic hepatitis in humans. Investigation of HEV replication is hampered by the lack of broadly applicable, efficient cell culture systems and tools for site-directed mutagenesis of HEV. The cell culture-adapted genotype 3c strain 47832c, which represents a typical genotype predominantly detected in Europe, has previously been used for several basic and applied research studies. Here, a plasmid-based reverse genetics system was developed for this strain, which efficiently rescued the infectious virus without the need for in vitro RNA transcription. The cotransfection of T7 RNA polymerase-expressing BSR/T7 cells with one plasmid encoding the full-length viral genome and two helper plasmids encoding vaccinia virus capping enzymes resulted in the production of infectious HEV, which could be serially passaged on A549/D3 cells. The parental and recombinant virus exhibited similar replication kinetics. A single point mutation creating an additional restriction enzyme site could be successfully introduced into the virus genome of progeny virus, indicating that the system is suitable for site-directed mutagenesis. This system is the first plasmid-based HEV reverse genetics system, as well as the first reverse genetics system for HEV genotype 3c, and should therefore be of broad use for basic and applied HEV research.

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Production of infectious dromedary camel hepatitis E virus by a reverse genetic system: Potential for zoonotic infection

Journal of Hepatology ◽

10.1016/j.jhep.2016.07.013 ◽

2016 ◽

Vol 65 (6) ◽

pp. 1104-1111 ◽

Cited By ~ 28

Author(s):

Tian-Cheng Li ◽

Xianfeng Zhou ◽

Sayaka Yoshizaki ◽

Yasushi Ami ◽

Yuriko Suzaki ◽

...

Keyword(s):

Hepatitis E Virus ◽

Genetic System ◽

Hepatitis E ◽

Reverse Genetic System ◽

Dromedary Camel ◽

Reverse Genetic ◽

Zoonotic Infection

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Mechanism of Cross-Species Transmission, Adaptive Evolution and Pathogenesis of Hepatitis E Virus

Viruses ◽

10.3390/v13050909 ◽

2021 ◽

Vol 13 (5) ◽

pp. 909

Author(s):

Putu Prathiwi Primadharsini ◽

Shigeo Nagashima ◽

Hiroaki Okamoto

Keyword(s):

Adaptive Evolution ◽

Hepatitis E Virus ◽

Current Knowledge ◽

Hepatitis E ◽

Oral Route ◽

Open Reading Frames ◽

Industrialized Countries ◽

Zoonotic Infection ◽

Host Determinants ◽

Viral Determinants

Hepatitis E virus (HEV) is the leading cause of acute hepatitis worldwide. While the transmission in developing countries is dominated by fecal-oral route via drinking contaminated water, the zoonotic transmission is the major route of HEV infection in industrialized countries. The discovery of new HEV strains in a growing number of animal species poses a risk to zoonotic infection. However, the exact mechanism and the determinant factors of zoonotic infection are not completely understood. This review will discuss the current knowledge on the mechanism of cross-species transmission of HEV infection, including viral determinants, such as the open reading frames (ORFs), codon usage and adaptive evolution, as well as host determinants, such as host cellular factors and the host immune status, which possibly play pivotal roles during this event. The pathogenesis of hepatitis E infection will be briefly discussed, including the special forms of this disease, including extrahepatic manifestations, chronic infection, and fulminant hepatitis in pregnant women.

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Review of Hepatitis E Virus in Rats: Evident Risk of Species Orthohepevirus C to Human Zoonotic Infection and Disease

Viruses ◽

10.3390/v12101148 ◽

2020 ◽

Vol 12 (10) ◽

pp. 1148

Author(s):

Gábor Reuter ◽

Ákos Boros ◽

Péter Pankovics

Keyword(s):

Hepatitis E Virus ◽

Current Knowledge ◽

Laboratory Diagnosis ◽

Hepatitis E ◽

Clinical Aspects ◽

Special Focus ◽

Human Pathogens ◽

Extrahepatic Manifestations ◽

Zoonotic Infection ◽

Symptomatic Disease

Hepatitis E virus (HEV) (family Hepeviridae) is one of the most common human pathogens, causing acute hepatitis and an increasingly recognized etiological agent in chronic hepatitis and extrahepatic manifestations. Recent studies reported that not only are the classical members of the species Orthohepevirus A (HEV-A) pathogenic to humans but a genetically highly divergent rat origin hepevirus (HEV-C1) in species Orthohepevirus C (HEV-C) is also able to cause zoonotic infection and symptomatic disease (hepatitis) in humans. This review summarizes the current knowledge of hepeviruses in rodents with special focus of rat origin HEV-C1. Cross-species transmission and genetic diversity of HEV-C1 and confirmation of HEV-C1 infections and symptomatic disease in humans re-opened the long-lasting and full of surprises story of HEV in human. This novel knowledge has a consequence to the epidemiology, clinical aspects, laboratory diagnosis, and prevention of HEV infection in humans.

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