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猪肺泡巨噬细胞感染猪繁殖与呼吸综合征病毒后的 miRNA 靶向转录组。

The miRNA-targeted transcriptome of porcine alveolar macrophages upon infection with Porcine Reproductive and Respiratory Syndrome Virus.

机构信息

GABI, INRA, AgroParisTech, Université Paris Saclay, Jouy-en-Josas, 78350, France.

Department of Population Health and Pathobiology, College of Veterinary Medicine, North Carolina State University, Raleigh, NC, United States.

出版信息

Sci Rep. 2019 Feb 28;9(1):3160. doi: 10.1038/s41598-019-39220-3.

Abstract

Host miRNAs are known to modulate the cell response to virus infections. We characterized the miRNA-targeted transcriptome of porcine alveolar macrophages (PAMs) at early times after infection with a subtype 1.1 strain of PRRSV (Porcine Reproductive and Respiratory Syndrome Virus). We performed the immunoprecipitation of RISC (RNA-induced Silencing Complex) followed by microarray analysis of the RISC-bound miRNA targets (RIP-Chip) to evaluate the relative enrichment or depletion of expressed genes in RISC. The miRNA-mediated regulation occurred early after PRRSV infection and decreased fast (1,241 and 141 RISC-bound genes at 7 h and 10 h post-infection, respectively); it affected several cell functions with evidence of miRNA buffering of upregulated interferon-related genes. Eight miRNAs were highly enriched in RISC of both control and infected cells with no evidence of differential expression. Although miR-335-5p was the miRNA with most predicted targets among enriched RISC-bound genes, no effects on surface markers, cytokine expression and PRRSV replication were detected upon miR-335-5p mimics of primary PAMs. Our results do not point to specific miRNA-driven mechanisms regulating the early response to infection with this PRRSV 1.1 strain and indicate that the miRNome expressed by steady-state PAMs reacts promptly to counterbalance PRRSV infection by a pervasive modulation of host functions.

摘要

宿主 microRNAs 被认为可调节细胞对病毒感染的反应。我们在感染猪繁殖与呼吸综合征病毒(PRRSV)1.1 亚型后早期,对猪肺泡巨噬细胞(PAMs)的 microRNA 靶向转录组进行了特征描述。我们进行了 RISC(RNA 诱导沉默复合物)的免疫沉淀,然后对 RISC 结合的 microRNA 靶标(RIP-Chip)进行了微阵列分析,以评估 RISC 中表达基因的相对富集或耗竭。microRNA 介导的调控发生在 PRRSV 感染后早期,且快速下降(感染后 7 h 和 10 h 时,分别有 1241 个和 141 个 RISC 结合基因;它影响了几种细胞功能,表明 microRNA 缓冲了上调的干扰素相关基因)。8 种 microRNA 在对照和感染细胞的 RISC 中高度富集,但没有差异表达的证据。尽管 miR-335-5p 在富集的 RISC 结合基因中具有最多的预测靶标,但在原发性 PAMs 中转录 miR-335-5p 模拟物时,并未检测到表面标志物、细胞因子表达和 PRRSV 复制的变化。我们的结果并未指向特定的 microRNA 驱动机制来调节对这种 PRRSV 1.1 株感染的早期反应,而是表明稳态 PAMs 表达的 microRNA 组通过对宿主功能的普遍调节,迅速做出反应以抵消 PRRSV 感染。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/532e/6395673/3fb6777db2ec/41598_2019_39220_Fig1_HTML.jpg

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