PEDV enters cells through clathrin-, caveolae-, and lipid raft-mediated endocytosis and traffics via the endo-/lysosome pathway

Abstract

With the emergence of highly pathogenic variant strains, porcine epidemic diarrhea virus (PEDV) has led to significant economic loss in the global swine industry. Many studies have described how coronaviruses enter cells, but information on PEDV invasion strategies remains insufficient. Given that the differences in gene sequences and pathogenicity between classical and mutant strains of PEDV may lead to diverse invasion mechanisms, this study focused on the cellular entry pathways and cellular transport of the PEDV GI and GII subtype strains in Vero cells and IPEC-J2 cells. We first characterized the kinetics of PEDV entry into cells and found that the highest invasion rate of PEDV was approximately 33% in the IPEC-J2 cells and approximately 100% in the Vero cells. To clarify the specific endocytic pathways, systematic research methods were used and showed that PEDV enters cells via the clathrin-and caveolae-mediated endocytosis pathways, in which dynamin II, clathrin heavy chain, Eps15, cholesterol, and caveolin-1 were indispensably involved. In addition, lipid raft extraction assay showed that PEDV can also enter cells through lipid raft-mediated endocytosis. To investigate the trafficking of internalized PEDV, we found that PEDV entry into cells relied on low pH and internalized virions reached lysosomes through the early endosome-late endosome-lysosome pathway. The results concretely revealed the entry mechanisms of PEDV and provided an insightful theoretical basis for the further understanding of PEDV pathogenesis and guidance for new targets of antiviral drugs.© The Author(s) 2020. This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article' s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article'

Antiviral activity of chlorpromazine, fluphenazine, perphenazine, prochlorperazine, and thioridazine towards RNA-viruses. A review

“…The summarized concentrations of the analyzed drugs inhibiting RNA viruses replication were shown in Table 3 . Moreover, the drug inhibits PEDV ( Wei et al, 2020 ) invasion. Noteworthy, the antiviral activity of chlorpromazine towards HCV is very promising as the obtained by Chamoun-Emanuelli et al (2013) results (IC 50 = 1.47 ± 0.32 μM and CC 50 /IC 50 = 6) ( Chamoun-Emanuelli et al, 2013 ) since both values are lower or very near of the toxic human plasma concentration.…”

“… Wei et al (2020) analyzed the impact of chlorpromazine on the infection of alphacoronavirus - porcine epidemic diarrhea virus (PEDV). The cytotoxicity results showed that the concentration of 30 μM and 50 μM did not affect the viability of the Vero and the IPEC-J2 cells, respectively.…”

“…The qRT-PCR results showed a significant decrease of PEDV invasion after chlorpromazine treatment (10 and 30 μM for Vero cells as well as 30 and 50 μM for IPEC-J2 cells). Moreover, chlorpromazine pre-treatment (to prevent clathrin assembly and further block clathrin-mediated endocytosis) and siRNA-mediated knockdown of clathrin heavy chain as well as EPS 15 (the critical component of clathrin-coated pits by interacting with adaptor protein 2) significantly reduce the invasion rate of PEDV into cells ( Wei et al, 2020 ).…”

Dynamic Dissection of Dynein and Kinesin-1 Cooperatively Mediated Intercellular Transport of Porcine Epidemic Diarrhea Coronavirus along Microtubule Using Single Virus Tracking

“…To understand PEDV outbreaks, various studies have been reported especially on the receptors [9,10] and endocytic pathways [11,12]. However, little is known about how PEDV particles traverse across the cytoplasm to facilitate their infection.…”

A Call for Action to Safely Deliver Oral Health Care during and Post COVID-19 Pandemic

“…In the initial stages of infection, cholesterol-rich lipid rafts play a fundamental role in viral entry into the cell [24]. These membrane microdomains facilitate both the interaction between the viral spike protein and the cellular ACE2 receptor [25] and the endocytosis process that takes place in the early stages of internalization of coronaviruses [26][27][28].…”