Porcine epidemic diarrhea virus infects and replicates in porcine alveolar macrophages

Park, Jung Eun; Shin, Hyun‐Jin

doi:10.1016/j.virusres.2014.07.038

Cited by 46 publications

(41 citation statements)

References 20 publications

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“…Here, we have developed a platform to facilitate the in vitro culture of PEDV by engineering Vero E6 cells to permanently express pAPN as a substrate for PEDV propagation. This modified cell line can augment PEDV replication, based on the observation that the majority of PEDV-positive intestinal samples used in our study gave rise to progressive syncytium formation in VeroE6-APN but not in parental VeroE6 cells (data not shown Recovery of PEDV using a full-length infectious clone been shown that growth of PEDV in these cells is markedly slower than in Vero cells (Cong et al, 2015;Park & Shin, 2014). Moreover, whilst ST cells permanently expressing high levels of pAPN (ST-pAPN-H/Q cells) could be efficiently infected by PEDV, the plaque size of PEDV-infected ST-pAPN-H/Q cells is apparently smaller than that of infected Vero cells (Nam & Lee, 2010).…”

Section: Discussionmentioning

confidence: 99%

Genetic manipulation of porcine epidemic diarrhoea virus recovered from a full-length infectious cDNA clone

Jengarn

Wongthida

Wanasen

et al. 2015

Journal of General Virology

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Porcine epidemic diarrhoea virus (PEDV) causes acute diarrhoea and dehydration in swine of all ages, with significant mortality in neonatal pigs. The recent rise of PEDV outbreaks in Asia and North America warrants an urgent search for effective vaccines. However, PEDV vaccine research has been hampered by difficulties in isolating and propagating the virus in mammalian cells, thereby complicating the recovery of infectious PEDV using a full-length infectious clone. Here, we engineered VeroE6 cells to stably express porcine aminopeptidase N (pAPN) and used them as a platform to obtain a high-growth variant of PEDV, termed PEDV AVCT12 . Subsequently, the full-length cDNA clone was constructed by assembling contiguous cDNA fragments encompassing the complete genome of PEDV AVCT12 in a bacterial artificial chromosome. Infectious PEDV could be recovered, and the rescued virus displayed phenotypic properties identical to the parental virus. Interestingly, we found that PEDV AVCT12 contained a C-terminal deletion of the spike gene, resulting in disruption of the ORF3 start codon. When a functional ORF3 gene was restored, the recombinant virus could not be rescued, suggesting that ORF3 could suppress PEDV replication in vitro. In addition, a high-growth and genetically stable recombinant PEDV expressing a foreign protein could be rescued by replacing the ORF3 gene with the mCherry gene. Together, the results of this study provide a means to generate genetically defined PEDV as a promising vaccine candidate.

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Section: Discussionmentioning

confidence: 99%

Genetic manipulation of porcine epidemic diarrhoea virus recovered from a full-length infectious cDNA clone

Jengarn

Wongthida

Wanasen

et al. 2015

Journal of General Virology

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“…In earlier studies, PEDV antigens were not detected by immunostaining within the spleen, liver or lungs of CV777‐infected pigs (Debouck et al., ) or in lungs of PEDV PC21A‐infected pigs (Jung et al., ), but the sensitivity of the RT‐qPCR assays used here is likely higher than the sensitivity of the immunostaining procedures. It has been reported (Park and Shin, ) that PEDV can replicate in pulmonary macrophages in vivo and thus extra‐intestinal replication of PEDV seems possible (Jung and Saif, ). The results presented here indicate that the US PEDV produced slightly more severe clinical signs than the cell culture‐grown Br1/87 strain.…”

Section: Discussionmentioning

confidence: 99%

Experimental Infection of Young Pigs with an Early European Strain of Porcine Epidemic Diarrhoea Virus and a Recent US Strain

Lohse

Krog

Strandbygaard

et al. 2016

Transbound Emerg Dis

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Outbreaks of porcine epidemic diarrhoea (PED) were reported across Europe during the 1980s and 1990s, but only sporadic outbreaks occurred in recent years. PED virus (PEDV) spread for the first time into the USA in 2013 and has caused severe economic losses. Retrospectively, it was found that two different strains of PEDV have been introduced into the United States, both are closely related to strains circulating in China where a new wave of the disease occurred from 2010 onwards. Since autumn 2014, new outbreaks of PED have occurred in Europe. In this study, weaned piglets were inoculated with an early European isolate (Br1/87) or faecal/intestinal suspensions derived from pigs infected with a recent European strain of PEDV (from Germany) or a US strain of PEDV. No evidence for infection resulted from inoculation of pigs with the German sample that contained high levels of PEDV RNA; there were no clinical signs, excretion of viral RNA or anti-PEDV antibody production. In contrast, all the pigs in the other two groups showed evidence of infection. Mild clinical signs of disease, mainly diarrhoea, occurred in piglets inoculated with the Br1/87 and US PEDV strains. PEDV RNA was detected throughout the intestine in euthanized animals at 4 days post-inoculation. In addition, within these animals, low levels of viral RNA were detected in lungs and livers with higher levels in spleens. Seroconversion against PEDV occurred in all surviving infected animals within 10 days. PEDV RNA excretion occurred for at least 2 weeks. The US PEDV RNA was detected at low levels in serum samples on multiple days. It is apparent that current diagnostic systems can detect infection by the different virus strains.

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“…Although macrophages infiltrated in the lamina propria also contained viral antigen as revealed by immunohistochemistry staining (Lee et al, 2000), it remains to be determined whether PEDV actually productively infected the intestinal macrophages. Porcine alveolar macrophages have been shown to support PEDV replication (Park and Shin, 2014). One recent study suggests that PEDV only replicates in Mo-DC prior to 24 h after infection, but not at later time points, as determined by virus titration of culture supernatants (Gao et al, 2015).…”

Section: Discussionmentioning

confidence: 99%

Porcine epidemic diarrhea virus does not replicate in porcine monocyte-derived dendritic cells, but activates the transcription of type I interferon and chemokine

Wang

Ohnstad

Nelsen

et al. 2017

Veterinary Microbiology

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Porcine epidemic diarrhea virus (PEDV) belongs to the alphacoronavirus of the Coronaviridae. It is the major etiological agent of the recent outbreaks of piglet diarrhea and death in the US. Limited knowledge is currently available regarding the role of dendritic cells in PEDV infection. Here, we observed that PEDV did not replicate in monocyte-derived dendritic cells as evidenced by the decrease of viral gene transcript copies in infected cells by qRT-PCR and the absence of viral proteins by immunofluorescence staining as well as the absence of virus particles in infected cells by transmission electron microscopy. In addition, PEDV did not compromise cell viability at 48, 72, and 96h after infection at either a MOI of 2.5 or 5. Interestingly, an increased transcription of type I interferon including interferon-α and β was observed in infected cells compared to mock infected cells. Surprisingly, we did not detect any interferon-β in the supernatants of infected cells. A slight increase in interferon-α protein production in the supernatants of PEDV-infected cells was observed compared to mock infected cells. We also observed a markedly increased transcription of interferon inducible protein -10 (IP-10). Overall, PEDV does not replicate in porcine Mo-DC, but activates the transcription of type I interferon and chemokine IP-10.

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Porcine epidemic diarrhea virus infects and replicates in porcine alveolar macrophages

Cited by 46 publications

References 20 publications

Genetic manipulation of porcine epidemic diarrhoea virus recovered from a full-length infectious cDNA clone

Genetic manipulation of porcine epidemic diarrhoea virus recovered from a full-length infectious cDNA clone

Experimental Infection of Young Pigs with an Early European Strain of Porcine Epidemic Diarrhoea Virus and a Recent US Strain

Porcine epidemic diarrhea virus does not replicate in porcine monocyte-derived dendritic cells, but activates the transcription of type I interferon and chemokine

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