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Aminopeptidase-N-independent entry of porcine epidemic diarrhea virus into Vero or porcine small intestine epithelial cells.
Abstract A monkey cell line Vero (ATCC CCL-81) is commonly used for porcine epidemic diarrhea virus (PEDV) propagation in vitro. However, it is still controversial whether the porcine aminopeptidase N (pAPN) counterpart on Vero cells (Vero-APN) confers PEDV entry. We found that endogenous expression of Vero-APN was undetectable in the mRNA and the protein levels in Vero cells. We cloned the partial Vero-APN gene (3340-bp) containing exons 1 to 9 from cellular DNA and subsequently generated two APN-knockout Vero cell lines by CRISPR/Cas9 approach. PEDV infection of two APN-knockout Vero cells had the same efficienc...
Source: Virology - March 1, 2018 Category: Virology Authors: Ji CM, Wang B, Zhou J, Huang YW Tags: Virology Source Type: research

Metalloprotease ADAM17 regulates porcine epidemic diarrhea virus infection by modifying aminopeptidase N.
Abstract Porcine epidemic diarrhea virus (PEDV) is a causative agent of porcine epidemic diarrhea (PED). PED, characterized by acute diarrhea, vomiting, dehydration, has caused serious economic losses in pig industry worldwide. Here, we demonstrate that activation of a disintergrin and metalloprotease 17 (ADAM17) induced the decrease of PEDV infection in HEK293 and IPEC-J2 cells and the downregulation of cell surface aminopeptidase N (APN) expression, an important entry factor for PEDV infection. Furthermore, overexpression of ADAM17 suppressed PEDV infection in HEK293 and IPEC-J2 cells, whereas ablation of ADAM17...
Source: Virology - February 20, 2018 Category: Virology Authors: Zhang J, Guo L, Yang L, Xu J, Zhang L, Feng L, Chen H, Wang Y Tags: Virology Source Type: research

Porcine aminopeptidase N mediated polarized infection by porcine epidemic diarrhea virus in target cells.
Abstract Infection of polarized intestinal epithelial cells by porcine epidemic diarrhea virus (PEDV) was characterized. Indirect immunofluorescence assay, real-time PCR, and transmission electron microscopy confirmed PEDV can be successfully propagated in immortalized swine small intestine epithelial cells (IECs). Infection involved porcine aminpeptidase N (pAPN), a reported cellular receptor for PEDV, transient expression of pAPN and siRNA targeted pAPN increased and decreased the infectivity of PEDV in IECs, respectively. Subsequently, polarized entry into and release from both Vero E6 and IECs was analyzed. PE...
Source: Virology - February 11, 2015 Category: Virology Authors: Cong Y, Li X, Bai Y, Lv X, Herrler G, Enjuanes L, Zhou X, Qu B, Meng F, Cong C, Ren X, Li G Tags: Virology Source Type: research