Project description:This study investigated the immunological function of PCV2 ORF5 by ectopic expression of PCV2 ORF5 in PK15 cell line. Identifying the functional role of each PCV2 ORF associated with host cell modulation may provide better knowledge about the pathogenesis of postweaning multisystemic wasting syndrome (PMWS). PCV2 ORF5 is recently identified and the functional role of ORF5 during the pathogenesis after PCV2 infection is largely unknown.
Project description:We employed deep sequencing technology to uncover cellular miRNAs differentially regulated after expression of each of three PCV2-encoded open reading frames (ORFs) in porcine kidney epithelial PK15 cells. Control PK15 vs. PCV2 ORF1, ORF2, ORF3-expressing PK15.
Project description:Based on the coinfection model of porcine circovirus type 2 (PCV2) and classical swine fever virus (CSFV), iTRAQ with LC-MS/MS was applied to map the cellular proteome profiles and explore cellular responses of cells coinfected with PCV2 and CSFV.
Project description:Purpose: Post-weaning multisystemic wasting syndrome (PMWS) is a disease associated with porcine circovirus type 2 (PCV2) infection. One main feature of PMWS is seriously decreased lymphocyte in lymphoid tissues. This research aims to investigate the molecular mechanism of evident lymphocyte depletion in spleen from Yorkshire x Landrace (YL) pigs after infection with PCV2. Methods: At the age of 6 weeks, 15 piglets of Laiwu (LW) and YL pigs were assigned randomly into two groups, respectively: 10 infected pigs and 5 uninfected pigs. Spleen tissues of each group were collected at 35 days post infection with PCV2. Three PCV2-infected YL pigs with apparently pathologic characteristics and mock-infected YL pigs were selected for RNA sequencing, respectively. The RNA extraction, library construction and RNA-Seq were performed on the Illumina HiSeq2500 at Beijing BioMarker Technologies (Beijing, China).Differential expression genes (DEGs) were filtrated from the result of transcriptome. The quantitative real-time PCR was performed to validate the expression patterns of candidate DEGs between LW and YL pigs. Results: The clean reads percolated from the raw reads were mapped to Sus scrofa genome (Sscrofa10.2) using Tophat2 software. The efficiency of alignment was about 76% between the six samples and reference genome, and approximate 50% clean reads were perfectly matched to pig genome. Two samples with high R-squared value (T1-T3: 0.98; T5-T6: 0.97) from PCV2 and mock-infected YL pigs, respectively, was performed differential expression analysis through DESeq after biologic repetition correlation detection. With the criterion of log FC (fold change) > 1.5 and of P value < 0.05, 90 significantly differentially expressed genes were identified between the two groups, of which 57 were up-regulated and 33 were down-regulated. Eleven candidate DEGs were confirmed by quantitative real-time PCR, which expression pattern were generally in accordance with the results of RNA-seq. Conclusions: Many DEGs participate in immune response and some of them are involved in cells proliferation or apoptosis, such as CD81, HGF, CXCL13, KLF11, PSMD4, CYCS and ACTC1. The expression changes of these DEGs may cause the lymphocyte depletion in spleen after challenge with PCV2.
Project description:We employed deep sequencing technology to uncover cellular miRNAs differentially regulated after expression of each of three PCV2-encoded open reading frames (ORFs) in porcine kidney epithelial PK15 cells.
Project description:Several recombinat viruses of porcine circovirus type 2 (PCV2),including P1, P2, ZJ-R, VL258, and VL264, have been found. The PK15 cells were transfected by the molecular clones of the abovementioned viruses, where specific sets of genes are up-regulated or down-regulated. We used microarrays to detail the global programme of gene expression and identified distinct functions of viruses or viral proteins. PK15 cells were selected at 12 hours post-transfection for RNA extraction and hybridization on Affymetrix microarrays. We sought to obtain the gene expression patterns of PK15 cells transfected with different molecular clones of the viruses.
Project description:Porcine circovirus type 2 (PCV2) can cause severe disease in the affected pigs, resulting in massive economic loss for the swine industry. Transcriptomics and proteomics approaches have been widely employed to identify the underlying molecular mechanisms of PCV2 infection. Numerous differentially expressed mRNAs, miRNAs, and proteins, together with their associated signaling pathways, have been identified, paving the way to further validation of their biological functions. Long non-coding RNA (lncRNA) is an important regulator of multiple biological processes. However, rare information regarding its role in the PCV2 infection has been obtained. Hence, in our study, RNA-seq was performed by infecting PK-15 cells with PCV2. Analysis of the differentially expressed genes (DEGs) suggested that cytoskeleton, apoptosis, cell division, and protein phosphorylation were significantly disturbed. Then, using stringent parameters, six lncRNAs were identified. Additionally, the potential targets of the lncRNAs were predicted using both cis- and trans-prediction methods. Interestingly, we found that the HOXB (Homeobox B) gene cluster was probably the target of the lncRNA LOC106505099. Enrichment analysis of the target genes showed that numerous developmental processes were altered during PCV2 infection. Therefore, our study revealed that lncRNAs could affect porcine embryo development through the regulation of the HOXB genes.
Project description:To further study of pcv2-related diseases, we have employed whole genome microarray expression profiling as a discovery platform to identify ileal differentially expressed genes of piglets after pcv2 infection. Infected and uninfected piglets were sampled and analyzed by whole genome microarray expression profiling. The result showed 43603 differencially expressed genes in ileum after PCV2 infection. Expression of 11 genes (IL-1α, IL-1β, IL-6, IFNε, C5, C1QA, CCL4, CCL5, CCL25, CXCL9, CD163) from this signature was quantified in the same RNA samples by real-time PCR, confirming low variability between samples.