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首页> 外文期刊>Journal of Medical Microbiology: An Official Journal of the Pathological Society of Great Britain and Ireland >Actinobacillus pleuropneumoniae culture supernatant antiviral effect against porcine reproductive and respiratory syndrome virus occurs prior to the viral genome replication and transcription through actin depolymerization
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Actinobacillus pleuropneumoniae culture supernatant antiviral effect against porcine reproductive and respiratory syndrome virus occurs prior to the viral genome replication and transcription through actin depolymerization

机译：胸膜肺炎放线杆菌培养上清液对猪繁殖与呼吸综合征病毒的抗病毒作用发生在病毒基因组通过肌动蛋白解聚复制和转录之前

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Purpose. Recently, the strong antiviral activity of an Actinobacillus pleuropneumoniae (App) culture supernatant against porcine reproductive and respiratory syndrome virus (PRRSV) was discovered. Following this finding, the objective of the present study was to understand how the App culture supernatant inhibits PRRSV replication in its natural targeted host cells, i.e. porcine alveolar macrophages (PAMs). Methodology. Several assays were conducted with App culture supernatant-treated PRRSV-infected cell lines, such as PAM, St-Jude porcine lung and MARC-145 cells. RT-qPCR assays were used to determine the expression levels of type I and II IFN mRNAs, viral genomic (gRNA) and sub-genomic RNAs (sgRNAs). Proteomic, Western blot and immunofluorescence assays were conducted to determine the involvement of actin filaments in the App culture supernatant antiviral effect. Results/Key findings. Type I and II IFN mRNA expressions were not upregulated by the App culture supernatant. Time courses of gRNA and sgRNA expression levels demonstrated that the App culture supernatant inhibits PRRSV infection before the first viral transcription cycle. Western blot experiments confirmed an increase in the expression of cofilin (actin cytoskeleton dynamics regulator) and immunofluorescence also demonstrated a significant decrease of actin filaments in App culture supernatant-treated PRRSV-infected PAM cells. App culture supernatant antiviral activity was also demonstrated against other PRRSV strains of genotypes I and II. Conclusion. App culture supernatant antiviral effect against PRRSV takes place early during PRRSV infection. Results suggest that App culture supernatant antiviral effect may take place via the activation of cofilin, which induces actin depolymerization and subsequently, probably affects PRRSV endocytosis. Other experiments are needed to fully validate this latest hypothesis.
机译：目的。最近，发现了胸膜肺炎放线杆菌（App）培养上清液对猪繁殖与呼吸综合征病毒（PRRSV）的强抗病毒活性。根据这一发现，本研究的目的是了解App培养上清液如何抑制PRRSV在其天然靶向宿主细胞即猪肺泡巨噬细胞（PAM）中的复制。方法。用App培养上清液处理的PRRSV感染的细胞系（如PAM，St-Jude猪肺和MARC-145细胞）进行了几种测定。 RT-qPCR分析用于确定I型和II型IFN mRNA，病毒基因组（gRNA）和亚基因组RNA（sgRNA）的表达水平。进行了蛋白质组学，蛋白质印迹和免疫荧光测定，以确定肌动蛋白丝参与App培养上清液的抗病毒作用。结果/主要发现。 App培养上清液未上调I型和II型IFN mRNA表达。 gRNA和sgRNA表达水平的时间过程证明，App培养上清液可在第一个病毒转录周期之前抑制PRRSV感染。 Western印迹实验证实了cofilin（肌动蛋白细胞骨架动力学调节剂）表达的增加，免疫荧光也证明了App培养上清液处理的PRRSV感染的PAM细胞中肌动蛋白丝的显着减少。还证明了应用培养上清液对其他PRRSV基因型I和II的抗病毒活性。结论。在PRRSV感染期间，应用培养上清液对PRRSV的抗病毒作用发生在早期。结果表明，App培养上清液的抗病毒作用可能是通过激活cofilin来实现的，cofilin的激活可引起肌动蛋白解聚，进而影响PRRSV的内吞作用。需要其他实验来完全验证这一最新假设。

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