Construction of an infectious cDNA clone of genotype 1 avian hepatitis E virus: characterization of its pathogenicity in broiler breeders and demonstration of its utility in studying the role of the hypervariable region in virus replication

Park, Soo-Jeong; Lee, Byung-Woo; Moon, Hyun-Woo; Sung, Haan Woo; Meng, Xiang‐Jin; Kwon, Hyun-Jung

doi:10.1099/vir.0.000045

Cited by 12 publications

(8 citation statements)

References 43 publications

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“…The clinical presentation caused by the virus can be variable. Broiler breeders which have been inoculated with avian HEV genotype 1, typically show more gross ovarian and microscopic lesions than younger chickens inoculated with genotype 2, while avian HEV genotype 3 infection of laying hens can exhibit BLS or HSS, but shows decreased egg production without gross ovary and microscopic lesions (Billam et al, 2005; Park et al, 2015; Zhao et al, 2017). There are several other diseases that could be caused by avian HEV, which are listed as follows (Table 1).…”

Section: Clinical Presentationmentioning

confidence: 99%

Avian Hepatitis E Virus: With the Trend of Genotypes and Host Expansion

Sun

Lin

et al. 2019

Front. Microbiol.

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Avian hepatitis E virus (HEV) is a single-stranded, positive-sense RNA virus with a complete genome of approximately 6.6 kb in size. To date, four major genotypes of avian HEV have been identified and classified into the Orthohepevirus B genus of the family Hepeviridae . The avian HEV associated with hepatitis-splenomegaly syndrome, big liver and spleen disease or hepatic rupture hemorrhage syndrome in chickens is genetically and antigenically related to mammalian HEV. With the increased genotypes of avian HEV identified, a broader host tropism is also notable in the epidemiological studies. Due to the lack of an efficient cell culture system, the mechanisms of avian HEV replication and pathogenesis are still poorly understood. The recent identification and characterization of animal strains of avian HEV has demonstrated the virus’ ability of cross-species infection. Although it has not yet been detected in humans, the potential threat of a zoonotic HEV capable of transmission to humans needs to be taken into consideration. This review article focuses on the current knowledge regarding avian HEV in virology, epidemiology, pathogenesis, clinical presentation, transmission, diagnosis and prevention. HIGHLIGHTS - The mechanisms of avian HEV replication and pathogenesis are still poorly understood due to the lack of an efficient cell culture system. - A broader host tropism is also notable in the epidemiological studies with the increased genotypes of avian HEV identified. - The recent identification and characterization of animal strains of avian HEV has demonstrated the virus’ ability of cross-species infection. - The potential threat of a zoonotic HEV capable of transmission to humans needs to be taken into consideration.

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Section: Clinical Presentationmentioning

confidence: 99%

Avian Hepatitis E Virus: With the Trend of Genotypes and Host Expansion

Sun

Lin

et al. 2019

Front. Microbiol.

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“…Avian HEV attachment to and infection of differently treated cell lines. Although there are no highly effective cell culture systems for avian HEV infection, the virus can attach to and infect LMH cells at low levels (32,57). The relationship between avian HEV infection and OATP1A2 expression levels was also evaluated in the differently treated LMH cell lines; however, the amount of avian HEV was detected only by TaqMan real-time RT-PCR.…”

Section: Primermentioning

confidence: 99%

Chicken Organic Anion-Transporting Polypeptide 1A2, a Novel Avian Hepatitis E Virus (HEV) ORF2-Interacting Protein, Is Involved in Avian HEV Infection

Fan

Liu

et al. 2019

J Virol

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Avian hepatitis E virus (HEV) is the main causative agent of big liver and spleen disease in chickens. Due to the absence of a highly effective cell culture system, there are few reports about the interaction between avian HEV and host cells. In this study, organic anion-transporting polypeptide 1A2 (OATP1A2) from chicken liver cells was identified to interact with ap237, a truncated avian HEV capsid protein spanning amino acids 313 to 549, by a glutathione S-transferase (GST) pulldown assay. GST pulldown and indirect enzyme-linked immunosorbent assays (ELISAs) further confirmed that the extracellular domain of OATP1A2 directly binds with ap237. The expression levels of OATP1A2 in host cells are positively correlated with the amounts of ap237 attachment and virus infection. The distribution of OATP1A2 in different tissues is consistent with avian HEV infection in vivo. Finally, when the functions of OATP1A2 in cells are inhibited by its substrates or an inhibitor or blocked by ap237 or anti-OATP1A2 sera, attachment to and infection of host cells by avian HEV are significantly reduced. Collectively, these results displayed for the first time that OATP1A2 interacts with the avian HEV capsid protein and can influence viral infection in host cells. The present study provides new insight to understand the process of avian HEV infection of host cells. IMPORTANCE The process of viral infection is centered around the interaction between the virus and host cells. Due to the lack of a highly effective cell culture system in vitro, there is little understanding about the interaction between avian HEV and its host cells. In this study, a total of seven host proteins were screened in chicken liver cells by a truncated avian HEV capsid protein (ap237) in which the host protein OATP1A2 interacted with ap237. Overexpression of OATP1A2 in the cells can promote ap237 adsorption as well as avian HEV adsorption and infection of the cells. When the function of OATP1A2 in cells was inhibited by substrates or inhibitors, attachment and infection by avian HEV significantly decreased. The distribution of OATP1A2 in different chicken tissues corresponded with that in tissues during avian HEV infection. This is the first finding that OATP1A2 is involved in viral infection of host cells.

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“…In previous reports, several infectious cDNA clones of avian HEV have already been constructed and proven to be a useful tool for in vivo studies [ 17 – 19 ]. In addition to that, the development and application of a detection system for negative-strand viral RNA provided valuable information about virus replication sites in vivo [ 20 ].…”

Section: Introductionmentioning

confidence: 99%

C-Terminal Amino Acids 471-507 of Avian Hepatitis E Virus Capsid Protein Are Crucial for Binding to Avian and Human Cells

et al. 2016

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The infection of chickens with avian Hepatitis E virus (avian HEV) can be asymptomatic or induces clinical signs characterized by increased mortality and decreased egg production in adult birds. Due to the lack of an efficient cell culture system for avian HEV, the interaction between virus and host cells is still barely understood. In this study, four truncated avian HEV capsid proteins (ORF2-1 – ORF2-4) with an identical 338aa deletion at the N-terminus and gradual deletions from 0, 42, 99 and 136aa at the C-terminus, respectively, were expressed and used to map the possible binding site within avian HEV capsid protein. Results from the binding assay showed that three truncated capsid proteins attached to avian LMH cells, but did not penetrate into cells. However, the shortest construct, ORF2-4, lost the capability of binding to cells suggesting that the presence of amino acids 471 to 507 of the capsid protein is crucial for the attachment. The construct ORF2-3 (aa339-507) was used to study the potential binding of avian HEV capsid protein to human and other avian species. It could be demonstrated that ORF2-3 was capable of binding to QT-35 cells from Japanese quail and human HepG2 cells but failed to bind to P815 cells. Additionally, chicken serum raised against ORF2-3 successfully blocked the binding to LMH cells. Treatment with heparin sodium salt or sodium chlorate significantly reduced binding of ORF2-3 to LMH cells. However, heparinase II treatment of LMH cells had no effect on binding of the ORF2-3 construct, suggesting a possible distinct attachment mechanism of avian as compared to human HEV. For the first time, interactions between avian HEV capsid protein and host cells were investigated demonstrating that aa471 to 507 of the capsid protein are needed to facilitate interaction with different kind of cells from different species.

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Construction of an infectious cDNA clone of genotype 1 avian hepatitis E virus: characterization of its pathogenicity in broiler breeders and demonstration of its utility in studying the role of the hypervariable region in virus replication

Cited by 12 publications

References 43 publications

Avian Hepatitis E Virus: With the Trend of Genotypes and Host Expansion

Avian Hepatitis E Virus: With the Trend of Genotypes and Host Expansion

Chicken Organic Anion-Transporting Polypeptide 1A2, a Novel Avian Hepatitis E Virus (HEV) ORF2-Interacting Protein, Is Involved in Avian HEV Infection

C-Terminal Amino Acids 471-507 of Avian Hepatitis E Virus Capsid Protein Are Crucial for Binding to Avian and Human Cells

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