Inhibition of porcine reproductive and respiratory syndrome virus replication by adenovirus-mediated RNA interference both in porcine alveolar macrophages and swine
Inhibition of porcine reproductive and respiratory syndrome virus replication by adenovirus-mediated RNA interference both in porcine alveolar macrophages and swine
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DOI：
10.1016/j.antiviral.2009.02.202
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发表时间：
2009-06-01
影响因子：
7.6
通讯作者：
Zeshan, Basit
中科院分区：
文献类型：
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作者：
Li, Guangming;Jiang, Ping;Zeshan, Basit
猪繁殖与呼吸综合征病毒（PRRSV）是造成许多养猪区重大经济损失的主要原因。目前的疫苗接种策略和抗病毒药物只能提供有限的保护。因此，有必要开发一种新的抗病毒策略。本研究构建了两种针对PRRSV S1株ORF1b表达短发夹rna （short hairpin rna, shRNAs）的重组腺病毒，并测定了其对PRRSV复制的抑制作用。结果表明，重组腺病毒介导的shrna预处理可显著抑制PRRSV SI株感染的Marc-145细胞的病毒RNA和蛋白水平。一种重组腺病毒（rAd-P2）在蛋白和ORF1b mRNA水平上也能有效抑制高毒力PRRSV SY0608株在Marc-145细胞和猪肺泡巨噬细胞中的复制。抗病毒效果呈剂量依赖性，持续时间至少为96小时。将26周龄仔猪分为4组，每组5头仔猪。1组和2组分别肌内接种rAd-P2和模拟构建体rAd-mP2。24 h后，1组、2组和3组肌内注射SY0608菌株。第4组没有受到挑战，但用PBS作为模拟。结果表明，rAd-P2能有效抑制猪血清和肺组织中的PRRSV病毒载量。与rAd-mP2阴性和PRRSV对照组相比，接种rAd-P2的猪的临床症状和病理病变较轻。这些结果表明，腺病毒介导的shrna在体外和体内均能充分抑制PRRSV感染。重组腺病毒介导的RNAi可能是一种控制PRRSV感染的新工具。当然，rAd-P2的保护效果还需要在未来大量的猪群中得到验证。(C) 2009 Elsevier B.V.版权所有
Porcine reproductive and respiratory syndrome virus (PRRSV) has been mainly responsible for the heavy economic losses in many swine-producing regions. Current vaccination strategies and antiviral drugs provide only limited protection. Consequently, there is a need to develop a new antiviral strategy. In this study, two recombinant adenoviruses expressing short-hairpin RNAs (shRNAs) directed against ORF1b of PRRSV S1 strain were constructed and the inhibition of PRRSV replication was determined. The results showed that pretreatment with these shRNAs delivered by recombinant adenovirus could induce a significant inhibition of viral RNA and protein level in Marc-145 cells infected with PRRSV SI strains. One recombinant adenovirus (rAd-P2) was found to be also effective in inhibiting the replication of highly virulent PRRSV SY0608 strain in Marc-145 cells and porcine alveolar macrophages at both the protein and ORF1b mRNA level. The antiviral effect was dose-dependent and sustained for at least 96 h. Twenty 6-week old piglets were assigned to four groups each with five piglets. Groups I and 2 were inoculated intramuscularly with rAd-P2 and mock construct rAd-mP2 individually. After 24 h, groups 1, 2 and 3 were challenged intramuscularly with the SY0608 strain. Group 4 remained unchallenged but with PBS as mock. The results showed that the viral load of PRRSV in serum and lung tissue of swine was suppressed effectively by rAd-P2. The clinical signs and pathological lesions in the pigs inoculated with rAd-P2 were milder than those in rAd-mP2 negative and PRRSV control. These results indicated that shRNAs mediated by the adenovirus could inhibit PRRSV infection sufficiently in vitro as well as in vivo. RNAi mediated by recombinant adenovirus might be a potential new tool for controlling PRRSV infection. Of course, the protective efficiency of rAd-P2 should be made by using a large number of pigs in future. (C) 2009 Elsevier B.V. All rights reserved.