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

Mammarenaviruses are enveloped and segmented negative-stranded RNA viruses that comprise several pathogenic members associated with severe human hemorrhagic fevers. Tacaribe virus (TCRV) is the prototype for the New World group of mammarenaviruses and is not only naturally attenuated but also phylogenetically and antigenically related to all South American pathogenic mammarenaviruses, particularly the Junín virus (JUNV), which is the etiological agent of Argentinian hemorrhagic fever (AHF). Moreover, since TCRV protects guinea pigs and non-human primates from lethal challenges with pathogenic strains of JUNV, it has already been considered as a potential live-attenuated virus vaccine candidate against AHF. Here, we report the development of a reverse genetic system that relies on T7 polymerase-driven intracellular expression of the complementary copy (antigenome) of both viral S and L RNA segments. Using this approach, we successfully recovered recombinant TCRV (rTCRV) that displayed growth properties resembling those of authentic TCRV. We also generated a chimeric recombinant TCRV expressing the JUNV glycoproteins, which propagated similarly to wild-type rTCRV. Moreover, a controlled modification within the S RNA 5′ non-coding terminal sequence diminished rTCRV propagation in a cell-type dependent manner, giving rise to new perspectives where the incorporation of additional attenuation markers could contribute to develop safe rTCRV-based vaccines against pathogenic mammarenaviruses.

Download Full-text

Establishment of an efficient reverse genetic system of Mumps virus S79 from cloned DNA

World Journal of Pediatrics ◽

10.1007/s12519-019-00286-8 ◽

2019 ◽

Vol 15 (5) ◽

pp. 499-505 ◽

Cited By ~ 1

Author(s):

Duo Zhou ◽

Meng-Ying Zhu ◽

Yi-Long Wang ◽

Xiao-Qiang Hao ◽

Dong-Ming Zhou ◽

...

Keyword(s):

Genetic System ◽

Mumps Virus ◽

Reverse Genetic System ◽

Reverse Genetic

Download Full-text

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

Download Full-text

Characterization and reverse genetic establishment of cattle derived Akabane virus in China

BMC Veterinary Research ◽

10.1186/s12917-021-03054-x ◽

2021 ◽

Vol 17 (1) ◽

Author(s):

Dongjie Chen ◽

Di Wang ◽

Fang Wei ◽

Yufang Kong ◽

Junhua Deng ◽

...

Keyword(s):

Genetic System ◽

Reverse Genetic System ◽

Akabane Virus ◽

Reverse Genetic ◽

Wild Type ◽

T7 Promoter ◽

Cytopathic Effects ◽

The World ◽

L Segment ◽

Genome Manipulation

Abstract Background Akabane virus (AKAV) is an important insect-borne virus which is widely distributed throughout the world except the Europe and is considered as a great threat to herbivore health. Results An AKAV strain defined as TJ2016 was firstly isolated from the bovine sera in China in 2016. Sequence analysis of the S and M segments suggested that the isolated AKAV strain was closely related to the AKAV strains JaGAr39 and JaLAB39, which belonged to AKAV genogroup II. To further study the pathogenic mechanism of AKAV, the full-length cDNA clone of TJ2016 S, M, and L segment was constructed separately into the TVT7R plasmid at the downsteam of T7 promoter and named as TVT7R-S, TVT7R-M, and TVT7R-L, respectively. The above three plasmids were further transfected into the BSR-T7/5 cells simultaneously with a ratio of 1:1:1 to produce the rescued virus AKAV. Compared with the parental wild type AKAV (wtAKAV), the rescued virus (rAKAV) was proved to be with similar cytopathic effects (CPE), plaque sizes and growth kinetics in BHK-21 cells. Conclusion We successfully isolated a AKAV strain TJ2016 from the sera of cattle and established a reverse genetic platform for AKAV genome manipulation. The established reverse genetic system is also a powerful tool for further research on AKAV pathogenesis and even vaccine studies.

Download Full-text

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.

Download Full-text

Seroprevalence of Dromedary Camel HEV in Domestic and Imported Camels from Saudi Arabia

Viruses ◽

10.3390/v12050553 ◽

2020 ◽

Vol 12 (5) ◽

pp. 553

Author(s):

Sherif Aly El-Kafrawy ◽

Ahmed Mohamed Hassan ◽

Mai Mohamed El-Daly ◽

Ishtiaq Qadri ◽

Ahmed Majdi Tolah ◽

...

Keyword(s):

Saudi Arabia ◽

Hepatitis E Virus ◽

Hepatitis E ◽

Health Concern ◽

P Value ◽

Contaminated Water ◽

Dromedary Camel ◽

Serum Samples ◽

Major Health

Hepatitis E Virus (HEV) imposes a major health concern in areas with very poor sanitation in Africa and Asia. The pathogen is transmitted mainly through ingesting contaminated water or food, coming into contact with affected people, and blood transfusions. Very few reports including old reports are available on the prevalence of HEV in Saudi Arabia in humans and no reports exist on HEV prevalence in camels. Dromedary camel trade and farming are increasing in Saudi Arabia with importation occurring unidirectionally from Africa to Saudi Arabia. DcHEV transmission to humans has been reported in one case from the United Arab Emeritus (UAE). This instigated us to perform this investigation of the seroprevalence of HEV in imported and domestic camels in Saudi Arabia. Serum samples were collected from imported and domestic camels. DcHEV-Abs were detected in collected sera using ELISA. The prevalence of DcHEV in the collected samples was 23.1% with slightly lower prevalence in imported camels than domestic camels (22.4% vs. 25.4%, p value = 0.3). Gender was significantly associated with the prevalence of HEV in the collected camels (p value = 0.015) where males (31.6%) were more infected than females (13.4%). This study is the first study to investigate the prevalence of HEV in dromedary camels from Saudi Arabia. The high seroprevalence of DcHEV in dromedaries might indicate their role as a zoonotic reservoir for viral infection to humans. Future HEV seroprevalence studies in humans are needed to investigate the role of DcHEV in the Saudi human population.

Download Full-text