Next Generation Sequencing analysis of the global expression profiling in bone marrow derived macrophages response to porcine circovirus type 2.
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ABSTRACT: To know the influences of PCV2 infection on macrophage phenotype, we examined the expression profiling of global genes in PCV2 infected-BMDMs by RNA-seq. The results revealed that expression of 369 genes were greater than twofold in PCV2 infected-BMDMs relative to their expression levels in control cells. In addition, the expression of 201 genes were lower than in PCV2-infected macrophages compared to that of control cells . Among these genes, we found that PCV2 infection strikingly induced expression of a subset of M1-associated genes, whereas decreased expression of multiple M2-associated genes, indicating that PCV2 infection likely reprograms the macrophage transcription phenotype.
Project description:Porcine alveolar macrophages (PAMs) play impoartant role in innate immunity. Porcine circovirus type 2 (PCV2) has been identified as the causal agent of postweaning multisystemic wasting syndrome, an economically important multifactorial disease of the swine industry worldwide. We used microarrays to study the transcriptome of PAMs infection with PCV2. PAMs were collected by bronchoalveolar lavage from health piglets (free of PCV2, PRRSV, PRV, CSFV, PPV), and PAMs were cultured for 48 hours and inoculated with 5 moi of PCV2.
Project description:Atherosclerosis is accelerated under diabetic conditions in part due to altered macrophage metabolism and function. Identifying critical factors to restore metabolic alterations and promote resolution of inflammation remains an unmet goal. MicroRNAs (miRs) orchestrate multiple signaling events in macrophages and regulate inflammation, yet their therapeutic potential in diabetes-associated atherosclerosis remains poorly understood. miRNA profiling of aortic intimal lesions from Ldlr-/- mice on a high-fat sucrose containing (HFSC) diet for 12 weeks revealed low expression of miR-369-3p. miR-369-3p was also reduced in peripheral blood mononuclear cells (PBMCs) from diabetic patients with coronary artery disease. Cell-type expression profiling showed miR-369-3p enrichment in aortic macrophages. Treatment with oxLDL reduced miR-369-3p expression in-stimulation mouse bone marrow-derived macrophages (BMDMs) downregulated miR-369-3p. Overexpression of miR-369-3p restored the oxLDL-mediated decrease of OXPHOS in BMDMs. Mechanistically, we found that miR-369-3p targeted the succinate receptor (GPR91) to alleviate the oxLDL-induced activation of inflammasome signaling pathways. Therapeutic administration of miR-369-3p in HFSC-fed Ldlr-/- mice reduced GPR91 expression in lesional macrophages and the diabetes-mediated accelerated atherosclerosis, evident by a decrease in plaque size and in the accumulation of pro-inflammatory Ly6Chi macrophages. RNA-seq analyses showed more pro-resolving pathways in plaque macrophages from miR-369-3p treated mice, consistent with an increase in macrophage efferocytosis in lesions. These findings establish a therapeutic role for miR-369-3p in halting diabetes-associated atherosclerosis by regulating GPR91 and macrophage succinate metabolism.
Project description:Porcine alveolar macrophages (PAMs) play impoartant role in innate immunity. Porcine circovirus type 2 (PCV2) has been identified as the causal agent of postweaning multisystemic wasting syndrome, an economically important multifactorial disease of the swine industry worldwide. We used microarrays to study the transcriptome of PAMs infection with PCV2.
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.
Project description:This study aimed to characterize differences in gene expression in piglets inoculated with porcine circovirus type 2 (PCV2), the essential causative agent of postweaning multisystemic wasting syndrome (PMWS). Comparisons between control and PCV2-inoculated pigs were done at five different time points: 1, 2, 5, 8, and 29 days post-inoculation. Experiment Overall Design: Seven-day-old caesarean-derived, colostrum-deprived piglets were distributed into two groups: control (n=8) and inoculated with 105.2 TCID50 of the Burgos PCV2 isolate (n=16). One control and 3 inoculated pigs were necropsied on days 1, 2, 5, and 8 post-infection (p.i.), the remaining pigs (4 of each group) were necropsied on day 29 p.i.
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:This study aimed to characterize differences in gene expression in piglets inoculated with porcine circovirus type 2 (PCV2), the essential causative agent of postweaning multisystemic wasting syndrome (PMWS). Comparisons between control and PCV2-inoculated pigs were done at five different time points: 0, 7, 14, 21 and 29 days post-inoculation. Keywords: time course
Project description:This study aimed to characterize differences in gene expression in piglets inoculated with porcine circovirus type 2 (PCV2), the essential causative agent of postweaning multisystemic wasting syndrome (PMWS). Comparisons between control and PCV2-inoculated pigs were done at five different time points: 1, 2, 5, 8, and 29 days post-inoculation. Keywords: longitudinal
Project description:This study aimed to characterize differences in gene expression in piglets inoculated with porcine circovirus type 2 (PCV2), the essential causative agent of postweaning multisystemic wasting syndrome (PMWS). Comparisons between control and PCV2-inoculated pigs were done at five different time points: 0, 7, 14, 21 and 29 days post-inoculation. Keywords: time course Seven-day-old caesarean-derived, colostrum-deprived piglets were distributed into two groups: control (n=4) and inoculated with 105.2 TCID50 of the Burgos PCV2 isolate (n=4). Pigs were bled at 0, 7, 14, 21, and 29 days post-inoculation.
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.