Broad antiviral peptides against PRRSV based on novel linear epitopes on porcine CD163
Broad antiviral peptides against PRRSV based on novel linear epitopes on porcine CD163
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DOI：
10.1016/j.ijbiomac.2022.03.040
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发表时间：
2022-03-15
影响因子：
8.2
通讯作者：
He, Fang
中科院分区：
文献类型：
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作者：
Han, Guangwei;Yang, Haotian;He, Fang
PRRSV给全球养猪业造成重大经济损失，需要创新的抗病毒药物。猪清道夫受体CD163已被确定为猪繁殖与呼吸综合征病毒（PRRSV）感染的重要融合受体。本研究基于广泛中和的单克隆抗体，从pCD163中制备了抗PRRSV的新型抗病毒肽。杆状病毒pCD163的SRCR-5-9能有效结合8系和1系的prrsv，阻断PAMs的感染。制备了一批靶向SRCR-5-9的单克隆抗体并进行了鉴定。8H2和4H7通过破坏病毒对PAMs的附着来阻断PRRSV感染。这些抗体使病毒滴度平均降低100-1000倍，病毒拷贝数降低约10(4)倍。8H2和4H7的线性表位分别定位于pCD163的SRCR6 （1- 30aa）和PSTI（1-15aa）。SRCR6 NI1718KT和PST SS1314AA突变分别使8H2和4H7对相应区域的识别失效。从线性表位衍生的多肽对不同谱系的prrsv表现出广泛的剂量依赖性抑制作用，并进一步调节prrsv相关的NF-kappa B途径。总之，这些发现加深了对PRRSV与pCD163受体相互作用的理解，并提供了针对PRRSV的替代通用抗病毒策略。
PRRSV causes major economic losses to swine industry world-wide, which requires innovative antiviral agents. Porcine scavenger receptor CD163 has been identified as an essential fusion receptor for Porcine reproductive and respiratory Syndrome Virus (PRRSV) infection. In this study, novel antiviral peptides from pCD163 against PRRSV were developed based on broad neutralizing monoclonal antibodies. SRCR-5-9 of pCD163 from baculovirus efficiently binds to PRRSVs of lineage 8 and lineage 1, blocking infection in PAMs. A batch of monoclonal antibodies targeting SRCR-5-9 were generated and characterized. 8H2 and 4H7 block PRRSV infection by the disruption in viral attachment to PAMs. Virus titer reduced 100-1000 folds in average and the virus copy number decreased about 10(4) folds with these antibodies. Linear epitopes of 8H2 and 4H7 were individually localized in SRCR6 (1-30 aa) and PSTI(1-15aa) of pCD163. Mutations of SRCR6 NI1718KT and PST SS1314AA abolished the recognition of 8H2 and 4H7 to the corresponding region individually. Peptides derived from the linear epitopes displayed a broad inhibitory effect on PRRSVs of different lineages in a dose-dependent manner and further modulated PRRSV-related NF-kappa B pathway. In conclusion, these findings deepen the understanding in the interaction between PRRSV and pCD163 receptor and provide alternative universal antiviral strategies against PRRSV.