Enhanced Replication of Hepatitis E Virus Strain 47832c in an A549-Derived Subclonal Cell Line

Schemmerer, Mathias; Apelt, Silke; Trojnar, E.; Ulrich, Rainer G.; Wenzel, Jürgen J.; Johne, Reimar

doi:10.3390/v8100267

Cited by 47 publications

(38 citation statements)

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“…4) while MeT's, derived from Sar-55, Mex-14, ZJ-1 or Kernow-C1, had little, if any, effect. It is even more interesting because HEV 47832c strain is a cell culture-adapted strain (Schemmerer et al, 2016). A simple hypothesis would be that HEV adaption to cell culture requires effective blockade of the IFN-β signaling.…”

Section: Discussionmentioning

confidence: 99%

Methyltransferase of a cell culture-adapted hepatitis E inhibits the MDA5 receptor signaling pathway

2019

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Hepatitis E virus (HEV) is a causative agent of acute hepatitis and jaundice. The number of human infections is approximated to be over 20 million cases per year. The transmission is mainly via the fecal-oral route and contaminated water and food are considered to be a major source of infection. As a mouse model is not available, a recent development of a cell culture-adapted HEV strain (47832c) is considered as a very important tools for molecular analysis of HEV pathogenesis in cells. Previously, we demonstrated that HEV-encoded methyltransferase (MeT) encoded by the 47832c strain inhibits MDA5-and RIG-I-mediated activation of interferon β (IFN-β)promoter. Here, we report that MeT impairs the phosphorylation and activation of interferon regulatory factor 3 and the p65 subunit of NF-κB in a dose-dependent manner. In addition, the MeT encoded by the 47832c, but not that of HEV clinical or field isolates (SAR-55, Mex-14, KC-1, and ZJ-1), displays the inhibitory effect. A deeper understanding of MeTmediated suppression of IFN-β expression would provide basis of the cell culture adaptation of HEV.

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Section: Discussionmentioning

confidence: 99%

Methyltransferase of a cell culture-adapted hepatitis E inhibits the MDA5 receptor signaling pathway

2019

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“…Recently, successful replication of HEV-3c strain 47832c (GenBank accession No. KC618403), in A549/D3 cells was demonstrated by Johne et al (2016;Schemmerer et al, 2016).…”

Section: Introductionmentioning

confidence: 90%

Evaluation of High-Pressure Processing in Inactivation of the Hepatitis E Virus

et al. 2020

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Hepatitis E virus (HEV) causes acute hepatitis with approximately 20 million cases per year globally. Based on genetic diversity, HEV is classified into different genotypes, with genotype 3 (HEV-3) being most prevalent in Europe and North America. The transmission of HEV-3 has been shown to be zoonotic and mainly associated with the consumption of raw or undercooked pork products. Herein, we investigated the efficacy of high-pressure processing (HPP) in inactivation of HEV-3 using a cell culture system. HPP has been indicated as a promising non-thermal pathogen inactivation strategy for treatment of certain high-risk food commodities, without any noticeable changes in their nature. For this purpose, we treated HEV-3 in media with different conditions of HPP: 400 MPa for 1 and 5 min, as well as 600 MPa for 1 and 5 min, at ambient temperature. All four HPP treatments of HEV in media were observed to result in a 2-log reduction in HEV load, as determined by the amounts of extracellular HEV RNA produced at 14-day post-infection, using the A549/D3 cell culture system. However, application of the same treatments to artificially contaminated pork pâté resulted in 0.5 log reduction in viral load. These results indicate that the efficacy of HPP treatment in the inactivation of HEV-3 is matrix-dependent, and independent of maximum pressure between 400 and 600 MPa and hold time between 1 and 5 min. Based on the obtained results, although the HPP treatment of pork pâté reduces the HEV-3 load, it might not be sufficient to fully mitigate the risk.

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“…In order to determine the limit of quantification of the infectivity assay, cell culture-adapted HEV-3 strain 47832c at ten concentrations from 5×10 2 to 1×10 6 genome copies per well were used to infect A549/D3 cells in triplicate experiments (29, 30). The infected and control cells, which were not exposed to viral particles, were cultured for 14 days.…”

Section: Methodsmentioning

confidence: 99%

“…However, due to the lack of reliable infectivity assays, most HEV inactivation studies to date have been limited to using surrogate viruses (27, 28). Recently, successful replication of HEV-3c strain 47832c (GenBank accession # KC618403), in A549/D3 cells was demonstrated by Johne and coworkers (29, 30). This system has been employed to study the temperature sensitivity of HEV (30), demonstrating a potential for this system to be used in other HEV inactivation studies.…”

Section: Introductionmentioning

confidence: 99%

“…Recently, successful replication of HEV-3c strain 47832c (GenBank accession # KC618403), in A549/D3 cells was demonstrated by Johne and coworkers (29, 30). This system has been employed to study the temperature sensitivity of HEV (30), demonstrating a potential for this system to be used in other HEV inactivation studies. Herein, we describe the employment of this HEV infectivity assay to examine the effect of HPP treatment on HEV infectivity in both cell culture media and ready-to-eat pork pâté.…”

Section: Introductionmentioning

confidence: 99%

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Evaluation of High-Pressure Processing in Inactivation of the Hepatitis E Virus

Nasheri

Doctor

Chen

et al. 2019

Preprint

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13Hepatitis E virus (HEV) causes acute hepatitis with approximately 20 million cases per year 14 globally. While HEV is endemic in certain regions of Asia, Africa and South America, it is 15 considered an emerging foodborne pathogen in developed countries. Based on genetic diversity, 16 HEV is classified into different genotypes, with genotype 3 (HEV-3) being most prevalent in 17 Europe and North America. The transmission of HEV-3 has been shown to be zoonotic and 18 mainly associated with the consumption of raw or undercooked pork products. Herein, we 19 investigated the efficacy of high-pressure processing (HPP) in the inactivation of HEV-3 using a 20 cell culture system. HPP has been indicated as a promising nonthermal pathogen inactivation 21 strategy for treatment of certain high-risk food commodities, without any noticeable changes in 22 their nature. For this purpose, we treated HEV-3 in media as well as in artificially inoculated 23 pork pâté, with different conditions of HPP: 400 MPa for 1 and 5 minutes, as well as 600 MPa 24 for 1 and 5 minutes, at ambient temperature. In general, we observed approximately a 2-log 25 reduction in HEV load by HPP treatments in media; however, similar treatment in the pork pâté 26 resulted in a much lower reduction in viral load. Therefore, the efficacy of HPP treatment in the 27 inactivation of HEV-3 is matrix-dependent. 28Importance: HEV is an emerging foodborne pathogen in industrialized countries, and its 29 transmission is associated with the consumption of contaminated undercooked pork product. In 30 this work, we employed an infectivity assay to investigate the potential of high-pressure in 31 inactivation of HEV in media and ready-to-eat pork pâté. We demonstrated that the effect of 32 HPP on inactivation of HEV depends on the surrounding matrix. 33 PCR 35 36 HEV is a single-stranded RNA virus with positive polarity belonging to the Hepeviridae family 37 (1). The HEV genome has 3 open reading frames (ORFs): ORF1 encodes a long non-structural 38 polyprotein with multiple functions; ORF2 encodes the viral capsid protein; and ORF3 encodes a 39 small phosphoprotein with structural and non-structural functions (2), (3). The Hepeviridae 40 contain two genera Orthohepevirus and Piscihepevirus, which infect a wide range of vertebrate 41 hosts (4). Four genotypes (HEV-1, HEV-2, HEV-3 and HEV-4) of the species Orthohepevirus A 42 are associated with human illness. HEV-1 and HEV-2 are restricted to humans and are prevalent 43 in regions with poor water sanitation, such as the developing countries of Asia, Africa, South and 44Central America (5, 6). On the other hand, HEV-3 and HEV-4 are considered to be zoonotic 45 pathogens as they have a much wider range of mammalian hosts including, among others, 46 domestic and wild swine and ruminants (7-9). Hepatocytes have been identified as the primary 47 sites of HEV replication, but the virus can replicate in other tissues such as epithelial cells of the 48 small intestine, placenta, and muscle (10-13). 49Clinical manifest...

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Enhanced Replication of Hepatitis E Virus Strain 47832c in an A549-Derived Subclonal Cell Line

Cited by 47 publications

References 31 publications

Methyltransferase of a cell culture-adapted hepatitis E inhibits the MDA5 receptor signaling pathway

Methyltransferase of a cell culture-adapted hepatitis E inhibits the MDA5 receptor signaling pathway

Evaluation of High-Pressure Processing in Inactivation of the Hepatitis E Virus

Evaluation of High-Pressure Processing in Inactivation of the Hepatitis E Virus

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