细胞内表达的抗猪繁殖与呼吸综合征病毒非结构蛋白4纳米抗体可抑制病毒复制。

Intracellularly expressed nanobodies against non-structural protein 4 of porcine reproductive and respiratory syndrome virus inhibit virus replication.

作者信息

Liu Hongliang, Liang Chao, Duan Hong, Zhang Xiaobin, Wang Xiangpeng, Xiao Shuqi, Zhou En-Min

机构信息

Department of Preventive Veterinary Medicine, College of Veterinary Medicine, Northwest A&F University, Yangling, 712100, Shaanxi, China.

出版信息

Biotechnol Lett. 2016 Jul;38(7):1081-8. doi: 10.1007/s10529-016-2086-3. Epub 2016 Mar 24.

DOI:10.1007/s10529-016-2086-3

PMID:27010387

Abstract

OBJECTIVE

To isolate specific nanobodies to porcine reproductive and respiratory syndrome virus (PRRSV) non-structural protein 4 (Nsp4) and investigate their potential antiviral activities.

RESULTS

Three PRRSV Nsp4-specific nanobodies were isolated from a phage display library of the variable domains of camelid heavy chain-only antibodies. Nanobody genes were introduced into MARC-145 cells using lentivirus vectors to establish cell lines stably expressing nanobodies. These intracellularly expressed nanobodies were tested for interaction with PRRSV-encoded Nsp4 within PRRSV-infected MARC-145 cells. Nb41 and Nb43 intrabodies each potently inhibited PRRSV replication, protected MARC-145 cells from PRRSV-induced cytopathic effect and fully blocked PRRSV replication at an MOI of 0.001 or lower.

CONCLUSION

Intracellularly expressed Nb41 and Nb43 potently suppressed PRRSV replication in MARC-145 cells. Nanobodies hold great potential for development as novel antiviral treatments for PRRSV infection.

摘要

目的

分离针对猪繁殖与呼吸综合征病毒（PRRSV）非结构蛋白4（Nsp4）的特异性纳米抗体，并研究其潜在的抗病毒活性。

结果

从骆驼科仅重链抗体可变区的噬菌体展示文库中分离出三种PRRSV Nsp4特异性纳米抗体。使用慢病毒载体将纳米抗体基因导入MARC-145细胞，以建立稳定表达纳米抗体的细胞系。检测这些细胞内表达的纳米抗体与PRRSV感染的MARC-145细胞中PRRSV编码的Nsp4的相互作用。Nb41和Nb43细胞内抗体均有效抑制PRRSV复制，保护MARC-145细胞免受PRRSV诱导的细胞病变效应，并在感染复数（MOI）为0.001或更低时完全阻断PRRSV复制。