RNA-sequence analysis of primary alveolar macrophages after in vitro infection with porcine reproductive and respiratory syndrome virus strains of differing virulence.
RNA-sequence analysis of primary alveolar macrophages after in vitro infection with porcine reproductive and respiratory syndrome virus strains of differing virulence.
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DOI：
10.1371/journal.pone.0091918
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发表时间：
2014
期刊：
影响因子：
3.7
通讯作者：
Botti S
中科院分区：
文献类型：
--
作者：
Badaoui B;Rutigliano T;Anselmo A;Vanhee M;Nauwynck H;Giuffra E;Botti S
猪繁殖与呼吸综合征病毒（Porcine reproductive and respiratory syndrome virus，PRRSV）主要感染猪肺泡巨噬细胞（porcine alveolar macrophages，PAM），导致猪繁殖与呼吸综合征（porcine reproductive and respiratory syndrome，PRRS）。迄今为止，大多数对感染PRRSV的PAM的转录组学研究都使用了微阵列技术。在此，我们通过RNA-Seq研究了两种欧洲PRRSV毒株感染后12 h的PAM体外转录组，其特征在于低（莱利斯塔，LV）和高（Lena）毒力。基因的表达水平，异构体，替代转录起始位点（TSS）和差异启动子的使用揭示了一个复杂的模式的转录和转录后基因调控感染后的两个菌株。基因本体分析证实，用Lena和LV菌株感染PAM影响与先天免疫应答直接相关的信号传导途径，包括干扰素调节因子（IRF）、RIG 1样受体、TLR和PKR途径。结果证实，干扰素信号转导是至关重要的PAM感染过程中的转录调控。IFN-β1和IFN-αω，但不是IFN-α，在感染LV或Lena株后上调。两种菌株均观察到经典途径的下调，例如先天性和适应性免疫应答、细胞死亡和TLR 3/TLR 7信号传导之间的相互作用，但Lena引发的下调比LV更强。该分析有助于更好地了解PRRSV和PAM之间的相互作用，并概述了PAM对不同毒力水平毒株的反应差异，这可能会导致新的PRRSV控制策略的发展。
Porcine reproductive and respiratory syndrome virus (PRRSV) mainly infects porcine alveolar macrophages (PAMs), resulting in porcine reproductive and respiratory syndrome (PRRS) in pigs. Most of the transcriptomic studies on PAMs infected with PRRSV conducted thus far have made use of microarray technology. Here, we investigated the transcriptome of PAMs in vitro at 12 h post-infection with two European PRRSV strains characterized by low (Lelystad, LV) and high (Lena) virulence through RNA-Seq. The expression levels of genes, isoforms, alternative transcription start sites (TSS) and differential promoter usage revealed a complex pattern of transcriptional and post-transcriptional gene regulation upon infection with the two strains. Gene ontology analysis confirmed that infection of PAMs with both the Lena and LV strains affected signaling pathways directly linked to the innate immune response, including interferon regulatory factors (IRF), RIG1-like receptors, TLRs and PKR pathways. The results confirmed that interferon signaling is crucial for transcriptional regulation during PAM infection. IFN-β1 and IFN-αω, but not IFN-α, were up-regulated following infection with either the LV or Lena strain. The down-regulation of canonical pathways, such as the interplay between the innate and adaptive immune responses, cell death and TLR3/TLR7 signaling, was observed for both strains, but Lena triggered a stronger down-regulation than LV. This analysis contributes to a better understanding of the interactions between PRRSV and PAMs and outlines the differences in the responses of PAMs to strains with different levels of virulence, which may lead to the development of new PRRSV control strategies.
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