猪流行性腹泻病毒功能性受体的鉴定  

Porcine Aminopeptidase N is a Functional Receptor for the PEDV Coronavirus

作　　者：李宝贤[1] 马广鹏[1] 葛俊伟[1] 李一经[1] 

机构地区：[1]东北农业大学动物医学院,黑龙江哈尔滨150030

出　　处：《病毒学报》2009年第3期220-225,共6页Chinese Journal of Virology

基　　金：国家自然科学基金项目资助(编号:30671574);东北农业大学创新团队发展计划(CX2008)

摘　　要：为研究猪氨基肽酶(Porcine Aminopeptidase N,pAPN)是否作为猪流行性腹泻病毒(Porcine epidemic diarrhea virus,PEDV)的细胞感染受体,通过转染技术,使PEDV非容许性细胞MDCK表达pAPN,并用PEDV感染转染细胞。结果发现转染的MDCK细胞可以感染PEDV,并且该病毒可以在转染细胞中连续传代。免疫荧光法鉴定存在病毒抗原。进一步实验证实,抗pAPN血清可以抑制PEDV感染转染的MDCK细胞。这些结果展示转染的MDCK细胞、pAPN表达及PEDV病毒复制之间存在直接联系,证明pAPN是PEDV的细胞感染受体之一。Porcine epidemic diarrhea virus （PEDV） causes lethal diarrhea in piglets that leads to great economic losses in East Asia. It was reported that aminopeptidase N （APN） was the receptor for Transmissible gastroenteritis virus （TGEV）, Human coronavirus 229E （HCoV-229E） and Feline coronavirus （FeCoV） which all belonged to group I coronavirus including PEDV. It was also confirmed previously that porcine aminopeptidase N （pAPN） could bind to PEDV, and anti-pAPN antibodies could inhibit the combination. To investigate whether pAPN was a receptor for PEDV, we transfected MDCK cells with porcine aminopeptidase （pAPN） cDNA and this enabled non-susceptible cells to support PEDV replication and serial viral propagation. Moreover, the infection was blocked by antibodies against pAPN, implying the critical role of pAPN during virus entry. In addition, immunofluorescence assays for detection of pAPN and PEDV antigens, together with neutralization assays using antibodies against pAPN, further confirmed the correlation between pAPN expression and viral replication in pAPN-transfected MDCK cells. These results indicated that pAPN is a functional receptor for PEDV.

关 键 词：受体 猪氨基肽酶 猪流行性腹泻病毒 冠状病毒 

分 类 号：S852.65[农业科学—基础兽医学]