Porcine epidemic diarrhea virus uses cell-surface heparan sulfate as an attachment factor

Huan, Changchao; Wang, Yue; Ni, Bo; Wang, Rui; Huang, Li; Ren, Xiaofeng; Tong, Guangzhi; Chen, Ding; Fan, Hongjie; Mao, Xiang

doi:10.1007/s00705-015-2408-0

Cited by 45 publications

(41 citation statements)

References 42 publications

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“…Our data show that the occludin protein does not mediate the initial attachment of virions to target cells, suggesting that occludin appears not to interact directly with virus at the cell surface. These findings indicate that another attachment receptor(s), such as cell surface heparan sulfate (54), may serve to bind the viral particles and thus help to concentrate viruses on the cell surface.…”

Section: Discussionmentioning

confidence: 92%

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: 92%

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

Luo

Guo

Zhang

et al. 2017

J Virol

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“…MDCK, IPEC-J2 and three colonies of MDCK -pAbcb1 were used to measure exogenous expression of porcine P-gp. qRT-PCR and western blot were used according to published methods54, with different primers (Table 1) and antibodies (Mdr-1(Santa Cruz, Heidelberg, Germany), dilution 1:200).…”

Section: Methodsmentioning

confidence: 99%

Abcb1 in Pigs: Molecular cloning, tissues distribution, functional analysis, and its effect on pharmacokinetics of enrofloxacin

Guo

Huang

Hong-yu

et al. 2016

Sci Rep

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P-glycoprotein (P-gp) is one of the best-known ATP-dependent efflux transporters, contributing to differences in pharmacokinetics and drug-drug interactions. Until now, studies on pig P-gp have been scarce. In our studies, the full-length porcine P-gp cDNA was cloned and expressed in a Madin-Darby Canine Kidney (MDCK) cell line. P-gp expression was then determined in tissues and its role in the pharmacokinetics of oral enrofloxacin in pigs was studied. The coding region of pig Abcb1 gene was 3,861 bp, encoding 1,286 amino acid residues (Mw = 141,966). Phylogenetic analysis indicated a close evolutionary relationship between porcine P-gp and those of cow and sheep. Pig P-gp was successfully stably overexpressed in MDCK cells and had efflux activity for rhodamine 123, a substrate of P-gp. Tissue distribution analysis indicated that P-gp was highly expressed in brain capillaries, small intestine, and liver. In MDCK-pAbcb1 cells, enrofloxacin was transported by P-gp with net efflux ratio of 2.48 and the efflux function was blocked by P-gp inhibitor verapamil. High expression of P-gp in the small intestine could modify the pharmacokinetics of orally administrated enrofloxacin by increasing the Cmax, AUC and Ka, which was demonstrated using verapamil, an inhibitor of P-gp.

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“…PEDV-infected cells produce distinct cytopathic effects including cell fusion, multinucleated syncytia formation, vacuolation, and detachment due to apoptotic cell death (Kim and Lee, 2014). A recent study shows that heparan sulfate on the cell-surface may function as an attachment factor for PEDV (Huan et al, 2015). As with TGEV, PEDV also utilizes aminopeptidase N (APN; CD13) as a functional receptor.…”

Section: Aminopeptidase N (Cd13) As the Cellular Receptor And Viral Tmentioning

confidence: 99%

Immune evasion of porcine enteric coronaviruses and viral modulation of antiviral innate signaling

Zhang

Yoo

2016

Virus Research

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Porcine epidemic diarrhea virus (PEDV) and porcine deltacoronavirus (PDCoV) are emerged and reemerging viruses in pigs, and together with transmissible gastroenteritis virus (TGEV), pose significant economic concerns to the swine industry. These viruses infect epithelial cells of the small intestine and cause watery diarrhea, dehydration, and a high mortality in neonatal piglets. Type I interferons (IFN-α/β) are major antiviral cytokines forming host innate immunity, and in turn, these enteric coronaviruses have evolved to modulate the host innate immune signaling during infection. Accumulating evidence however suggests that IFN induction and signaling in the intestinal epithelial cells differ from other epithelial cells, largely due to distinct features of the gut epithelial mucosal surface and commensal microflora, and it appears that type III interferon (IFN-λ) plays a key role to maintain the antiviral state in the gut. This review describes the recent understanding on the immune evasion strategies of porcine enteric coronaviruses and the role of different types of IFNs for intestinal antiviral innate immunity.

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Porcine epidemic diarrhea virus uses cell-surface heparan sulfate as an attachment factor

Cited by 45 publications

References 42 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

Abcb1 in Pigs: Molecular cloning, tissues distribution, functional analysis, and its effect on pharmacokinetics of enrofloxacin

Immune evasion of porcine enteric coronaviruses and viral modulation of antiviral innate signaling

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