Putative phage-display epitopes of the porcine epidemic diarrhea virus S1 protein and their anti-viral activity

Cao, Liyan; Ge, Xianping; Gao, Yu; Zarlenga, Dante S.; Wang, Kexiong; Li, Xunliang; Qin, Zhaoheng; Yin, Xin; Liu, Jisheng; Ren, Xiaofeng; Guangxin, LI

doi:10.1007/s11262-015-1234-5

Cited by 18 publications

(21 citation statements)

References 38 publications

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“…In addition, the relocalization of occludin upon PEDV infection is also seen in noninfected cells, suggesting indirect effects of PEDV on the cellular distribution of occludin. Since others have reported that PEDV-infected Vero E6 and IPEC-J2 cells do not produce type I interferon (38,39), we infer that the internalization of occludin in noninfected cells may be induced by some other pathophysiologic stimuli, such as proinflammatory cytokines (40). Jung and colleagues demonstrated that PEDV infection resulted in structurally altered tight junction proteins ZO-1 and E-cadherin in the villous epithelium in vivo (41).…”

Section: Discussionmentioning

confidence: 83%

Tight Junction Protein Occludin Is a Porcine Epidemic Diarrhea Virus Entry Factor

Luo

Guo

Zhang

et al. 2017

J Virol

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Porcine epidemic diarrhea virus (PEDV), the causative agent of porcine epidemic diarrhea, has caused huge economic losses in pig-producing countries. Although PEDV was long believed to replicate in the intestinal epithelium by using aminopeptidase N as a receptor, the mechanisms of PEDV infection are not fully characterized. In this study, we found that PEDV infection of epithelial cells results in disruption of the tight junctional distribution of occludin to its intracellular location. Overexpression of occludin in target cells makes them more susceptible to PEDV infection, whereas ablation of occludin expression by use of small interfering RNA (siRNA) in target cells significantly reduces their susceptibility to virus infection. However, the results observed with occludin siRNA indicate that occludin is not required for virus attachment. We conclude that occludin plays an essential role in PEDV infection at the postbinding stages. Furthermore, we observed that macropinocytosis inhibitors blocked occludin internalization and virus entry, indicating that virus entry and occludin internalization are closely coupled. However, the macropinocytosis inhibitors could not impede virus replication once the virus had entered host cells. This suggests that occludin internalization by macropinocytosis or a macropinocytosis-like process is involved in the virus entry events. Immunofluorescence confocal microscopy showed that PEDV was trapped at cellular junctional regions upon macropinocytosis inhibitor treatment, indicating that occludin may serve as a scaffold in the vicinity of virus entry. Collectively, these data show that occludin plays an essential role in PEDV infection during late entry events. Our observation may provide novel insights into PEDV infection and related pathogenesis.IMPORTANCE Tight junctions are highly specialized membrane domains whose main function is to attach adjacent cells to each other, thereby forming intercellular seals. Here we investigate, for the first time, the role of the tight junction protein occludin in PEDV infection. We observed that PEDV infection induced the internalization of occludin. By using genetic modification methods, we demonstrate that occludin plays an essential role in PEDV infection. Moreover, PEDV entry and occludin internalization seem to be closely coupled. Our findings reveal a new mechanism of PEDV infection.KEYWORDS PEDV, occludin, tight junction C oronaviruses are a group of positive-strand RNA viruses belonging to the family Coronaviridae in the order Nidovirales. They are broadly distributed among mammalian and avian species, and they cause acute and persistent infections. In most cases, coronaviruses are generally associated with significant respiratory and/or intestinal tract diseases (1, 2). Porcine epidemic diarrhea virus (PEDV) has been identified as the etiologic agent of porcine epidemic diarrhea (PED), and it causes diarrhea, vomiting, and dehydration in infected swine (3, 4). Outbreaks of PED have occurred frequently in

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Section: Discussionmentioning

confidence: 83%

Tight Junction Protein Occludin Is a Porcine Epidemic Diarrhea Virus Entry Factor

Luo

Guo

Zhang

et al. 2017

J Virol

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“…Three different groups of S1 proteins from co‐viral infections shared less than 12% amino acid sequence identities with each other (Wang, Deng et al., ). The B‐cell response is directed against the spike protein of coronaviruses (Cao et al., ) and it plays an important role in pathogenesis of virus infection.…”

Section: Discussionmentioning

confidence: 99%

The detection and phylogenetic analysis of porcine deltacoronavirus from Guangdong Province in Southern China

Mai

Feng

Chen

et al. 2017

Transbound Emerg Dis

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Porcine deltacoronavirus (PDCoV) is a newly discovered coronavirus that causes diarrhoea, vomiting and dehydration in sucking and nursing piglets. It was first reported in Hong Kong in 2012 and has since been discovered in the United States, Canada, South Korea, mainland China, Thailand and Laos. PDCoV has been experimentally proved to lead to diarrhoea in swine and it was detected positive in pigs in Guangdong, southern China. In our study, 252 faecal and intestinal samples from sucking piglets and sows with diarrhoea were surveyed for common enteric viruses. We found a prevalence of PDCoV (21.8%), porcine epidemic diarrhoea virus (65.5%), transmissible gastroenteritis virus (0%), rotavirus group A (25.0%) and porcine kobuvirus (68.7%). We isolated 13 PDCoV strains and discovered that PDCoV infections were often co-infections with kobuvirus rather than the commonly linked porcine epidemic diarrhoea virus. Phylogenetic analysis of S gene and N gene revealed that 11 of 13 PDCoV strains belonged to Chinese lineage. As for the left two strains, one single strain (CHN-GD16-05) belonged to American and Korean lineages while another strain (CHN-GD16-03) was similar to a Thai strain, but only in the S gene. This suggested a possible recombination event between the Thai and the newly described Chinese strain.

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“…Total RNA was extracted using RNA extraction kit (Qiagen, Germany), and cDNA was synthesized by using cDNA reverse transcription kit (ThermoFisher, USA) with oligo dt primer. The cDNA was then subjected to real-time PCR with primer pairs specific for PEDV and βactin (Cao et al 2015). The β-actin gene served as the endogenous control.…”

Section: Qds-605 Labeled Sdab-mc37 As Probes Imaging Pedv In Vero Cellsmentioning

confidence: 99%

Single-domain antibodies as promising experimental tools in imaging and isolation of porcine epidemic diarrhea virus

Yang

Yin

et al. 2018

Appl Microbiol Biotechnol

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Single-domain antibody (sdAb) or nanobody possesses specific features non-accessible for conventional antibodies that make them suitable for research and biotechnological applications. Porcine epidemic diarrhea virus (PEDV) causes lethal diarrhea in piglets, resulting in great economic losses all over the world. To detect and isolate PEDV rapidly and accurately is important for the control and further research of the clinical PEDV strains. In this study, four sdAb fragments (sdAb-Mc19/29/30/37) targeting the membrane (M) protein of PEDV were selected from sdAb library that was constructed through M protein-immunized Camelus bactrianus. The selected sdAb-Mcs were solubly expressed in Escherichia coli. The functional characteristics analysis revealed that the recombinant sdAb-Mcs have excellent binding activity and specificity to M protein but have no neutralizing activity to PEDV. For further application, sdAb-Mc37 was conjugated with quantum dots to synthesize a nanoprobe for imaging PEDV in vero cells. The observed fluorescence in vero cells clearly reflects that PEDV virions can be reliably recognized and labeled by the nanoprobe. Furthermore, the sdAb-Mc29 was conjugated with superparamagnetic nanobeads to construct immunomagnetic nanobeads (IMNBs) used to isolate PEDV. One PEDV strain was successfully isolated from clinical fecal sample, suggesting IMNBs as a novel and efficient tool suitable for PEDV isolation from clinical samples. This study provided a novel application and substantiated the suitability of sdAb as a specific binder for the isolation of viruses.

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Putative phage-display epitopes of the porcine epidemic diarrhea virus S1 protein and their anti-viral activity

Cited by 18 publications

References 38 publications

Tight Junction Protein Occludin Is a Porcine Epidemic Diarrhea Virus Entry Factor

Tight Junction Protein Occludin Is a Porcine Epidemic Diarrhea Virus Entry Factor

The detection and phylogenetic analysis of porcine deltacoronavirus from Guangdong Province in Southern China

Single-domain antibodies as promising experimental tools in imaging and isolation of porcine epidemic diarrhea virus

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