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Trypsin-induced hemagglutination activity of porcine epidemic diarrhea virus.

ABSTRACT: The hemagglutination (HA) activity of porcine epidemic diarrhea virus (PEDV) was investigated. Two cell-adapted strains of PEDV (KPEDV-9 and SM98LVec) were subjected to HA test against erythrocytes of various origin. Both strains showed HA activity with rabbit erythrocytes only after treatment with trypsin or neuraminidase. Optimal conditions for inducing HA activity of PEDV were 2 h incubation at 37 degrees C using phosphate-buffered saline containing 0.1% BSA. These results suggest that the HA activity of PEDV is most likely caused by proteolytic action on it, which could be developed as a new diagnostic method to rapidly detect and differentiate PEDV infections from other enteric diseases.

SUBMITTER: Park JE

PROVIDER: S-EPMC7087193 | biostudies-literature | 2010 Apr

REPOSITORIES: biostudies-literature

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Trypsin-induced hemagglutination activity of porcine epidemic diarrhea virus.

Park Jung-Eun JE Cruz Deu John M DJ Shin Hyun-Jin HJ

Archives of virology 20100309 4

The hemagglutination (HA) activity of porcine epidemic diarrhea virus (PEDV) was investigated. Two cell-adapted strains of PEDV (KPEDV-9 and SM98LVec) were subjected to HA test against erythrocytes of various origin. Both strains showed HA activity with rabbit erythrocytes only after treatment with trypsin or neuraminidase. Optimal conditions for inducing HA activity of PEDV were 2 h incubation at 37 degrees C using phosphate-buffered saline containing 0.1% BSA. These results suggest that the HA ...[more]

PMID: 20217154

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Project description:Background: Porcine Epidemic Diarrhea Virus (PEDV) is a coronavirus that infects the intestinal tract and causes diarrhea and vomiting in older pigs or extreme dehydration and death that could reach 100% mortality in neonatal piglets. In the US, the first PEDV outbreaks occurred in 2013 and since then US PEDV strains have quickly spread throughout the US and worldwide, causing significant economic and public health concerns. Currently two conditionally approved vaccines exist in the US, but there is no live attenuated vaccine, which is considered the best option in controlling PEDV by inducing transferrable mucosal immunity to susceptible neonatal piglets. In this study, we passaged an US PEDV isolate under various conditions to generate three strains and characterized their growth and antigenicity in cell culture using various assays including Western blot analysis, serum neutralization assay, sequencing analysis and confocal microscopy. Finally, these strains were evaluated for pathogenicity in nursing piglets (1-4 days old).Results: One of the PEDV strains generated in this study (designated as PEDV 8aa) is able to replicate in cells without any protease and grows to a high titer of >8 log10 TCID50/ml in cell culture. Interestingly, replication of PEDV 8aa was severely reduced by trypsin and this correlated with the inhibition of virus attachment and entry into the cells. In neonatal nursing piglets, PEDV 8aa (passage number 70 or 105) was found to be fully attenuated with limited virus shedding.Conclusions: These results suggest that applying selective pressure during viral passages can facilitate attainment of viral attenuation and that PEDV 8aa warrants further investigation as an attenuated vaccine.

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Project description:Porcine epidemic diarrhea virus (PEDV) invades porcine intestinal epithelial cells (IECs) and causes diarrhea and dehydration in pigs. In the present study, we showed a suppression of PEDV infection in porcine jejunum intestinal epithelial cells (IPEC-J2) by an increase in autophagy. Autophagy was activated by rapamycin at a dose that does not affect cell viability and tight junction permeability. The induction of autophagy was examined by LC3I/LC3II conversion. To confirm the autophagic-flux (entire autophagy pathway), autophagolysosomes were examined by an immunofluorescence assay. Pre-treatment with rapamycin significantly restricted not only a 1 h infection but also a longer infection (24 h) with PEDV, while this effect disappeared when autophagy was blocked. Co-localization of PEDV and autophagosomes suggests that PEDV could be a target of autophagy. Moreover, alleviation of PEDV-induced cell death in IPEC-J2 cells pretreated with rapamycin demonstrates a protective effect of rapamycin against PEDV-induced epithelial cell death. Collectively, the present study suggests an early prevention against PEDV infection in IPEC-J2 cells via autophagy that might be an effective strategy for the restriction of PEDV, and opens up the possibility of the use of rapamycin in vivo as an effective prophylactic and prevention treatment.

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