Next Generation Sequencing of miRNA in PRRSV-infected PAMs
Ontology highlight
ABSTRACT:
PROVIDER: PRJNA692737 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| SRR13450790.fastq.gz | Fastqsanger.gz | |||
| SRR13450791.fastq.gz | Fastqsanger.gz | |||
| SRR13450792.fastq.gz | Fastqsanger.gz | |||
| SRR13450793.fastq.gz | Fastqsanger.gz | |||
| SRR13450794.fastq.gz | Fastqsanger.gz |
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Project description:Porcine reproductive and respiratory syndrome (PRRSV) is a devastating pathogen for the pig industry worldwide. PRRSV mainly infects porcine alveolar macrophage (PAM). In the present study, single-cell RNA sequencing (scRNA-seq) was employed to characterize the host transcriptome responses of PAMs infected with a virulent PRRSV strain in vivo and ex vivo. Increase ofpro-inflammatory and anti-apoptotic genes was correlated with the increase of infection (expression of the virus ORF7 transcript).
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Project description:Porcine reproductive and respiratory syndrome (PRRS), which caused by the porcine reproductive and respiratory syndrome virus (PRRSV), is a serious viral disease affecting global swine industry. At present, PRRSV vaccines fail to prevent this disease. Consequently, new antiviral strategies to compensate for the inefficacy of available vaccines are urgently required. Lysine acetylation is an important post-translational modification (PTM) regulating an array of pathological and physiological conditions. In this study, we profiled the global acetylome using acetylation specific antibody based enrichment and Tandem mass tag (TMT) label LC-MS in PRRSV-infected pulmonary alveolar macrophages (PAMs). As a result, 3731 lysine acetylation sites on 1421 cellular proteins were identified and quantified 6 hours post infection (hpi). Bioinformatics analysis of the differentially acetylated proteins revealed their involvement in various biological processes, including the host immune response and energy metabolism.
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Project description:Transcriptome analysis of a PRRSV-infected and control PAMs
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Project description:Porcine reproductive and respiratory syndrome (PRRS) is an economically important disease with a significant impact on the pig industry. It is caused by PRRS virus (PRRSV), which predominantly infects and replicates in porcine pulmonary alveolar macrophages (PAMs), providing a useful in vitro model for understanding the host immune response to PRRSV infection. We pretreated PAMs with porcine IFN-α to induce an antiviral state within the cells and subsequently infected them with highly pathogenic (HP)-PRRSV. Changes in global gene expression in IFN-α-pretreated PAMs in response to HP-PRRSV infection were determined by RNA-sequence analysis. A total of 346 differentially expressed genes (DEGs) were transcriptionally upregulated by porcine IFN-α. Among these up-regulated DEGs, 93 showed significantly attenuated expression levels in response to HP-PRRSV infection. These attenuated DEGs were remarkably enriched in immune-response-related terms, such as 'RIG-Ilike receptor signaling pathway', 'Response to virus', and 'Response to stimulus'. Notably, expression levels of 93.8% (15/16) of antiviral genes (such as PKR, OAS1, IFIT1(ISG56) and ISG15), and genes encoding retinoic-acid-inducible gene-I protein (LOC100737466(DDX58), a cytoplasmic viral RNA sensor), interferon regulatory factor 7 (IRF7, a key transcriptional regulator of type I IFN-dependent immune response), signal transducer and activator of transcription 1 (LOC100738308(STAT1), a transcription factor for IFN-stimulated genes), and tumor necrosis factor-related apoptosis-inducing ligand (TNFSF10, a cytokine initiating proapoptotic signaling cascades) were significantly attenuated by HP-PRRSV infection. These results suggest that HP-PRRSV can counteract the type I IFN-induced antiviral state by interfering with the expression of genes involved in the host-defense response.
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Project description:Porcine reproductive and respiratory syndrome (PRRS), caused by PRRS virus (PRRSV), is the most economically important disease in pig populations. Lung damage is one major pathological condition following PRRSV infection, often leading to animal death. In vivo, PRRSV productive infection occurs predominately in alveolar macrophages of the lung. Here, transcriptome profiling of pulmonary alveolar macrophages (PAMs) from Tongcheng piglets pre- and post- infection of highly pathogenic PRRSV has been performed using porcine Affymetrix GeneChip. All animal procedures were performed according to protocols approved by the Biological Studies Animal Care and Use Committee of Hubei Province, China. Piglets used in this study were free from PRRSV, pseudorabies virus (PRV) and porcine circovirus type 2 (PCV2) determined by ELISA test for serum antibodies. Twelve of 5-week-old boars were obtained from three litters (four piglets per litter), and raised in pathogen-free facilities. In order to perform a paired experiment, every four full-sib individuals were divided equally into two groups: one infected group and one control group with 6 piglets in each group. The infected groups were challenged with PRRSV-Wuh2 (3 ml/15 kg, 10-5 TCID50/ml) by intramuscular inoculation. Slaughters were carried out at 0 days post-infection (dpi) for uninfected (control) groups, and at 5 or 7 dpi for infected groups. The PAMs for microarray analysis were collected by bronchoalveolar lavage from three uninfected pigs and three infected pigs at 5 dpi. Total of 6 microarrays have been hybridized in this experiment.
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