Application of RNAscope technology to studying the infection dynamics of a Chinese porcine epidemic diarrhea virus variant strain BJ2011C in neonatal piglets

Cai, Yu; Wang, Di; Zhou, Lei; Ge, Xinna; Guo, Xin; Han, Jun; Yang, Hanchun

doi:10.1016/j.vetmic.2019.07.003

Cited by 11 publications

(8 citation statements)

References 36 publications

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“…Building on the finding that PEDV infection influence iron levels via TfR1 mediated PEDV entry, we next investigated whether altering iron levels affected TfR1 mediated PEDV infection. As PEDV mainly infects the small intestine [ 27 , 28 ], we selected it as the site for further experiments. Fig 3A illustrates the in vivo treatments to the ligated small intestines of newborn piglets.…”

Section: Resultsmentioning

confidence: 99%

Transferrin receptor 1 levels at the cell surface influence the susceptibility of newborn piglets to PEDV infection

2020

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Porcine epidemic diarrhea virus (PEDV) mainly infects the intestinal epithelial cells of newborn piglets causing acute, severe atrophic enteritis. The underlying mechanisms of PEDV infection and the reasons why newborn piglets are more susceptible than older pigs remain incompletely understood. Iron deficiency is common in newborn piglets. Here we found that high levels of transferrin receptor 1 (TfR1) distributed in the apical tissue of the intestinal villi of newborns, and intracellular iron levels influence the susceptibility of newborn piglets to PEDV. We show that iron deficiency induced by deferoxamine (DFO, an iron chelating agent) promotes PEDV infection while iron accumulation induced by ferric ammonium citrate (FAC, an iron supplement) impairs PEDV infection in vitro and in vivo. Besides, PEDV infection was inhibited by occluding TfR1 with antibodies or decreasing TfR1 expression. Additionally, PEDV infection was increased in PEDV-resistant Caco-2 and HEK 293T cells over-expressed porcine TfR1. Mechanistically, the PEDV S1 protein interacts with the extracellular region of TfR1 during PEDV entry, promotes TfR1 re-localization and clustering, then activates TfR1 tyrosine phosphorylation mediated by Src kinase, and heightens the internalization of TfR1, thereby promoting PEDV entry. Taken together, these data suggest that the higher expression of TfR1 in the apical tissue of the intestinal villi caused by iron deficiency, accounts for newborn piglets being acutely susceptible to PEDV.

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Section: Resultsmentioning

confidence: 99%

Transferrin receptor 1 levels at the cell surface influence the susceptibility of newborn piglets to PEDV infection

2020

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“…These findings are consistent with a previous study (Ma et al, 2015) and suggest that PDCoV can replicate in the epithelium of the small intestine. In addition, to confirm the IHC results, RNAscope, a novel technology for RNA hybridization that can achieve sensitive detection of viral RNAs in situ at the single-cell level (Cai et al, 2019;Wang et al, 2012), was used to detect the distribution of PDCoV RNA in the sectioned tissues of PDCoV-infected piglets. Our results showed that positive signals were predominantly distributed in the cytoplasm of the villous epithelial cells of the small intestine, which are consistent with the IHC results.…”

Section: Histotropism Analysis Of Pdcov In Pigletsmentioning

confidence: 99%

A strain of porcine deltacoronavirus: Genomic characterization, pathogenicity and its full‐length cDNA infectious clone

Zhou

Zhang

et al. 2020

Transbound Emerg Dis

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As a novel enteropathogenic coronavirus, porcine deltacoronavirus (PDCoV) warrants further investigation. In this study, a Chinese PDCoV strain, designated CHN-HN-1601, was isolated from the faeces of a diarrhoeic piglet. After plaque purification, the genome was determined which shared 97.5%-99.5% nucleotide identities with 71 representative PDCoV strains available in the GenBank. The pathogenic properties of CHN-HN-1601 were evaluated using 5-day-old piglets. All inoculated piglets developed severe diarrhoea from 2 days post-infection (dpi) onwards. To our surprise, two periods of diarrhoea starting from 2 to 7 dpi and from 13 to 19 dpi were observed in affected piglets during the experiment. Faecal viral shedding of the inoculated piglets was detected by real-time RT-PCR, with viral shedding peaked at 4 and 16 dpi, respectively. At necropsy at 5 dpi, the main gross lesions included transparent, thin-walled and gas-distended intestines containing yellow watery contents.Further histopathological examinations, including haematoxylin and eosin staining, immunohistochemistry and RNAscope in situ hybridization, revealed that the virus infection caused severe villous atrophy of the small intestines, with PDCoV antigen and RNA mainly distributed in the cytoplasm of the villous epithelial cells of jejunum and ileum in piglets. The dynamic production of PDCoV-specific IgG and neutralizing antibodies in serum of the affected piglets was also assessed using a whole virus-based ELISA and an immunofluorescence assay-based neutralization test, respectively. Furthermore, a full-length cDNA infectious clone of CHN-HN-1601 was constructed using a bacterial artificial chromosome system. The rescued virus exhibited in vitro growth and pathogenic properties similar to the parental virus. Taken together, our study not only enriches the information of PDCoV, but also provides a useful reverse genetics platform for further pathogenesis exploration of the virus.

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“…Previous researches have recommended that the ambient meteorological factors and air pollutants were acted as the risk factors for acute to severe respiratory infection by carrying fatal micro pathogens and organisms. The micro pollutants and pathogens affect the human body by decreasing the level of immunity and more vulnerable and prone toward the SARS-CoV-2 (Cai et al, 2019;Hernandez et al, 2018;Pettersson et al, 2019).…”

Section: Introductionmentioning

confidence: 99%

Significance between Air pollutants, Meteorological Factors and COVID-19 Infections: Probable Evidences in India

Sahoo

2020

Preprint

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SARS-CoV-2 (Coronavirus) disease represents the causative agent with a potentially fatal risk which is having great global human health concern. Earlier studies suggested that air pollutants and meteorological factors were considered as the risk factors for acute respiratory infection, which carries harmful pathogens and affects the immunity. The study intended to explore the correlation between air pollutants, meteorological factors and the daily reported infection cases caused by novel coronavirus in India. The daily positive infected cases, air pollution and meteorological factors in 288 districts were collected from January 30, 2020 to April 23, 2020 in India. Speraman’s correlation and generalised additive model were applied to investigate the correlations of four air pollutants (PM2.5, PM10, NO2 and SO2) and eight meteorological factors (Temp, DTR, RH, AH, AP, RF, WS and WD) with COVID-19 infected cases. The study indicated that a 10 µg/m3 increase during (Lag0-14) in PM2.5, PM10 and NO2 was resulted in 2.21% (95%CI: 1.13 to 3.29), 2.67% (95% CI: 0.33 to 5.01) and 4.56 (95% CI: 2.22 to 6.90) increase in daily counts of COVID 19 infected cases respectively. However, only 1 unit increase in meteorological factor levels in case of daily mean temperature and DTR during (Lag0-14) associated with 3.78% (95%CI: 1.81 to 5.75) and 1.82% (95% CI: -1.74 to 5.38) rise of COVID-19 infected cases respectively. In addition, SO2 and relative humidity were negatively associated with COVID-19 infected cases at Lag0-14 with decrease of 7.23% (95% CI: -10.99 to -3.47) and 1.11% (95% CI: -3.45 to 1.23) for SO2 and for relative humidity respectively. The study recommended that there is significant relationship between air pollutants and meteorological factors with COVID-19 infected cases, which substantially explain the effect of national lockdown and suggested positive implications for control and prevention of the spread of SARS-CoV-2 disease.

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Application of RNAscope technology to studying the infection dynamics of a Chinese porcine epidemic diarrhea virus variant strain BJ2011C in neonatal piglets

Cited by 11 publications

References 36 publications

Transferrin receptor 1 levels at the cell surface influence the susceptibility of newborn piglets to PEDV infection

Transferrin receptor 1 levels at the cell surface influence the susceptibility of newborn piglets to PEDV infection

A strain of porcine deltacoronavirus: Genomic characterization, pathogenicity and its full‐length cDNA infectious clone

Significance between Air pollutants, Meteorological Factors and COVID-19 Infections: Probable Evidences in India

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