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猪氨肽酶N介导猪流行性腹泻病毒在靶细胞中的极化感染。

Porcine aminopeptidase N mediated polarized infection by porcine epidemic diarrhea virus in target cells.

作者信息

Cong Yingying, Li Xiaoxue, Bai Yunyun, Lv Xiaonan, Herrler Georg, Enjuanes Luis, Zhou Xingdong, Qu Bo, Meng Fandan, Cong Chengcheng, Ren Xiaofeng, Li Guangxing

机构信息

College of Veterinary Medicine, Northeast Agricultural University, Harbin 150030, China.

College of Veterinary Medicine, Northeast Agricultural University, Harbin 150030, China; CAS Key Lab for Biomedical Effects of Nanomaterials and Nanosafety, National Center for Nanoscience & Technology of China, Beijing 100090, China.

出版信息

Virology. 2015 Apr;478:1-8. doi: 10.1016/j.virol.2015.01.020. Epub 2015 Feb 11.

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. PEDV entry into polarized cells and pAPN grown on membrane inserts occurs via apical membrane. The progeny virus released into the medium was also quantified which demonstrated that PEDV is preferentially released from the apical membrane. Collectively, our data demonstrate that pAPN, the cellular receptor for PEDV, mediates polarized PEDV infection. These results imply the possibility that PEDV infection may proceed by lateral spread of virus in intestinal epithelial cells.

摘要

对猪流行性腹泻病毒(PEDV)感染极化肠道上皮细胞的特性进行了研究。间接免疫荧光测定、实时聚合酶链反应和透射电子显微镜证实PEDV能够在永生化猪小肠上皮细胞(IECs)中成功繁殖。感染涉及猪氨肽酶N(pAPN),它是一种已报道的PEDV细胞受体,pAPN的瞬时表达和靶向pAPN的小干扰RNA分别增加和降低了PEDV在IECs中的感染性。随后,分析了PEDV进入和离开Vero E6细胞和IECs的极化过程。PEDV通过顶膜进入极化细胞以及在膜插入物上生长的pAPN。还对释放到培养基中的子代病毒进行了定量,结果表明PEDV优先从顶膜释放。总的来说,我们的数据表明,PEDV的细胞受体pAPN介导了PEDV的极化感染。这些结果暗示了PEDV感染可能通过病毒在肠道上皮细胞中的侧向传播而发生的可能性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c274/7112081/c4ecdbf93633/gr1_lrg.jpg

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