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.

中文翻译：

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PEDV通过网格蛋白，海绵体和脂质筏介导的内吞作用进入细胞，并通过内体/溶酶体途径运输。

随着高致病性变异株的出现，猪流行性腹泻病毒（PEDV）已导致全球养猪业的重大经济损失。许多研究已经描述了冠状病毒如何进入细胞，但是关于PEDV入侵策略的信息仍然不足。鉴于PEDV经典株和突变株之间基因序列和致病性的差异可能导致多种入侵机制，本研究着重研究了Vero细胞和IPEC-J2细胞中PEDV GI和GII亚型菌株的细胞进入途径和细胞运输。 。我们首先表征了PEDV进入细胞的动力学，发现PEDV的最高侵袭率在IPEC-J2细胞中约为33％，在Vero细胞中约为100％。为了阐明特定的内吞途径，使用了系统的研究方法，结果表明PEDV通过网格蛋白和小窝蛋白介导的内吞途径进入细胞，其中动力蛋白II，网格蛋白重链，Eps15，胆固醇和小窝蛋白1必不可少。另外，脂筏提取试验表明，PEDV也可以通过脂筏介导的内吞作用进入细胞。为了研究内在化的PEDV的运输，我们发现PEDV依赖于低pH进入细胞，并且内化的病毒颗粒通过早期的内体-晚期内体-溶酶体途径到达了溶酶体。研究结果具体揭示了PEDV的进入机制，为进一步了解PEDV的发病机理和抗病毒药物新靶点的指导提供了深刻的理论基础。