Inhibition of highly pathogenic porcine reproductive and respiratory syndrome virus replication by recombinant pseudorabies virus-mediated RNA interference in piglets

被引:12
作者
Cao, Su-fang [1 ]
Guo, Qing-yong [2 ]
Wang, Yan [1 ]
机构
[1] Henan Univ Anim Husb Econ, Dept Vet Med, Mingli Rd, Zhengzhou 450046, Peoples R China
[2] Xinjiang Agr Univ, Coll Anim Sci, Urumqi 830052, Peoples R China
关键词
HP-PRRSV; Recombination pseudorabies virus; RNAi; siRNA; PROTECTIVE IMMUNITY; ADENOVIRAL VECTORS; DISEASE; PROTEIN; ESCAPE; IMMUNOGENICITY; CONSTRUCTION; SUPPRESSION; EFFICACY; SWINE;
D O I
10.1016/j.vetmic.2015.10.016
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
Highly pathogenic porcine reproductive and respiratory syndrome virus (HP-PRRSV) is a variant of porcine reproductive and respiratory syndrome virus (PRRSV) which, in recent years, has caused heavy economic losses to swine-producing areas. Although current vaccines are somewhat prophylactic, they provide only limited protection. Furthermore, there are currently no effective anti-HP-PRRSV drugs. Consequently, it is necessary to develop novel antiviral strategies. In the present study, three recombinant pseudorabies viruses (PRV) expressing siRNAs against the ORF7 of HP-PRRSV strain HN1 (PRV gG-/siRNAN1, PRV gG-/siRNAN2, and PRV gG-/siRNAN3) were evaluated for the inhibition of HP-PRRSV replication. The results indicated that recombinant PRV-mediated siRNA could significantly decrease the replication of traditional PRRSV strain H1 at mRNA and protein levels in Marc-145 cells. Moreover, one recombinant PRV (PRV gG-/siRNAN2) was found to be inhibit the multiplication of HP-PRRSV strain HN1 effectively in Marc-145 cells at both the protein and ORF7 mRNA level. Twenty 21-day old healthy weaned piglets were divided into four groups of five piglets each. Groups 1 and 2 were injected i.m. with PRV gG-/siRNAN2 and PRV gG-/siRNANeg individually. The piglets in group 3 were challenged with the HP-PRRSV control. After 24 h, the piglets in groups 1-3 were challenged i.m. with HP-PRRSV strain HN1, while those in group 4 were i.m. administered with PBS as a negative control. The results showed that HP-PRRSV in serum and lung samples from piglets was effectively inhibited by PRV gG-/siRNAN2. The clinical signs and gross lesions of piglets inoculated with PRV gG-/siRNAN2 were significantly less invasive than those of the PRV gG-/siRNANeg group and HP-PRRSV control group. These results showed that siRNAs mediated by recombinant PRV could effectively suppress HP-PRRSV replication in vitro as well as in vivo. RNAi mediated by recombinant PRV presents a potential novel method to prevent HP-PRRSV infections in swine. However, the protective efficiency of PRV gG-/siRNAN2 should be assessed in a larger number of piglets in future studies. Crown Copyright (C) 2015 Published by Elsevier B.V. All rights reserved.
引用
收藏
页码:212 / 220
页数:9
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