Project description:One hundred Hampshire by Duroc crossbred pigs (HD) and 100 NE Index line pigs (I) were infected with porcine reproductive and respiratory syndrome virus (PRRSV) and evaluated for resistance/susceptibility. Controls (100/line) were uninfected littermates to infected pigs. Viremia (V), weight change (WTΔ), and rectal temperature at 0, 4, 7, and 14 days post-infection (dpi) were recorded. Lung, bronchial lymph node (BLN), and blood tissue were collected at necropsy (14 dpi). Infected pigs were classified as low or high responders to PRRSV based on the first principal component (PC) from principal component analyses of all variables. Low responders to PRRSV (low PRRSV burden) and their uninfected littermates were assigned to low (L) class. High responders to PRRSV (high PRRSV burden) and their uninfected littermates were assigned to high (H) class. Infected pigs in the L-class had high WTΔ, low V, and few lung lesions; H-class pigs had low WTΔ, high V, and many lung lesions. RNA was extracted from lung and BLN tissue of the seven highest and seven lowest responders per line and from each of their control littermates. A control reference design was used and cDNA from each reference sample tissue was prepared from pooled RNA extracted from two control pigs from each line whose infected littermates had a PC value of 0. Design variables in data analyses were line (I vs HD), class (H vs L), treatment (infected vs uninfected controls), and slide/pig as error.
Project description:To analyze gene expression profiles at the single-cell level in pigs infected with PRRSV (Porcine Reproductive and Respiratory Syndrome Virus), aiming to understand the dynamic changes in gene expression, immune responses, and cellular interactions during PRRSV infection, and identify potential targets for intervention or prevention
Project description:The goal of this study was to produce a deep, global analysis of gene expression changes that occured following infection of normal porcine alveolar macrophages (PAMs) with PRRSV. The goal was to examine the gene expression changes to help determine the mechanisms that result in reduced function and immunosuppression observed in PRRSV-infected pigs. Keywords: time course of infection
Project description:The goal of this study was to produce a deep, global analysis of gene expression changes that occured following infection of normal porcine alveolar macrophages (PAMs) with PRRSV. The goal was to examine the gene expression changes to help determine the mechanisms that result in reduced function and immunosuppression observed in PRRSV-infected pigs. Keywords: time course of infection The PAMs were infected in culture at an MOI of 10 with PRRSV strains VR-2332 and incubated at 37C until 6, 12, 16 or 24 hours post infection. Total cellular RNA was collected from each at the appropriate time. SAGE libraries were prepared from each infected time point as well as from noninfected PAMs. The SAGE libraries were sequenced to at least 95,000 tags each.
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.
Project description:Porcine reproductive and respiratory syndrome (PRRS) is a highly contagious disease caused by porcine reproductive and respiratory syndrome virus (PRRSV). It clinically manifests as fever, bleeding, and respiratory symptoms in pigs of different age groups, as well as reproductive disorders, causing huge economic losses to the global swine industry. RNA interference (RNAi), a highly conserved mechanism mediating post-transcriptional gene silencing, exerts antiviral effects by cleaving viral double-stranded RNA to generate virus-derived siRNA (vsiRNA). VsiRNA-mediated antiviral immunity is widespread in plants and invertebrates, while in mammals, studies have focused on reducing PRRSV viral load in pigs via siRNA delivery. Therefore, PRRSV strains (LY and LY-R) were primarily selected as virus models to infect PAM and Marc-145 cells, respectively. Then, miRNA and vsiRNA analysis were performed and bioinformatics analysis revealed multiple differentially expressed miRNAs following PRRSV infection. Further enrichment analysis indicated that these miRNAs were mainly enriched in metabolic, endocytic, and phosphorylation signaling pathways. Additionally, in Marc-145 cells infected with PRRSV-LY, high abundance of vsiRNA was detected within the PRRSV-N genome. The N-vsiRNA was inserted into vector psilencer4.1. to construct a PS-N plasmid, which is capable of exerting anti-PRRSV (LY, LY-R, and NADC30-like) activity in Marc-145 cells. Finally, the PS-N plasmid was formulated with PLGA-PEI to prepare a nanoparticle mixture, which was delivered to pigs via intramuscular injection to alleviate tissue damage caused by PRRSV-LY infection to a certain extent. In summary, the porcine RNAi system and vsiRNA generated via viral cleavage play a critical role in inhibiting virus replication. This further demonstrates the feasibility of vsiRNA-based antiviral strategies and offers a reference for the prevention and control of various RNA viruses.
Project description:Porcine reproductive and respiratory syndrome (PRRS) is a highly contagious disease caused by porcine reproductive and respiratory syndrome virus (PRRSV). It clinically manifests as fever, bleeding, and respiratory symptoms in pigs of different age groups, as well as reproductive disorders, causing huge economic losses to the global swine industry. RNA interference (RNAi), a highly conserved mechanism mediating post-transcriptional gene silencing, exerts antiviral effects by cleaving viral double-stranded RNA to generate virus-derived siRNA (vsiRNA). VsiRNA-mediated antiviral immunity is widespread in plants and invertebrates, while in mammals, studies have focused on reducing PRRSV viral load in pigs via siRNA delivery. Therefore, PRRSV strains (LY and LY-R) were primarily selected as virus models to infect PAM and Marc-145 cells, respectively. Then, miRNA and vsiRNA analysis were performed and bioinformatics analysis revealed multiple differentially expressed miRNAs following PRRSV infection. Further enrichment analysis indicated that these miRNAs were mainly enriched in metabolic, endocytic, and phosphorylation signaling pathways. Additionally, in Marc-145 cells infected with PRRSV-LY, high abundance of vsiRNA was detected within the PRRSV-N genome. The N-vsiRNA was inserted into vector psilencer4.1. to construct a PS-N plasmid, which is capable of exerting anti-PRRSV (LY, LY-R, and NADC30-like) activity in Marc-145 cells. Finally, the PS-N plasmid was formulated with PLGA-PEI to prepare a nanoparticle mixture, which was delivered to pigs via intramuscular injection to alleviate tissue damage caused by PRRSV-LY infection to a certain extent. In summary, the porcine RNAi system and vsiRNA generated via viral cleavage play a critical role in inhibiting virus replication. This further demonstrates the feasibility of vsiRNA-based antiviral strategies and offers a reference for the prevention and control of various RNA viruses.
Project description:Porcine reproductive and respiratory disease (PRRS) is the most important disease in swine industry worldwide. However, strategies such as vaccination and good biosecurity are not consistently successful to eliminate PRRSV. Though some interesting pathways have been tentatively examined recently, host molecular pathways utilized by PRRSV and the protective immune responses in resistant to PRRSV are largely unknown. In order to answer these questions, we herein characterize changes in global gene expressions in multiple tissues [tonsil, tracheobronchial lymph nodes (TBLN), Cranial lung (CR Lung), and distal lung (D Lung)] in response to PRRSV of high and low virulence. Both vaccinated and unvaccinated pigs are used for this study. Based on Ingenuity Pathway Analysis (IPA), molecule bases of some “black boxes” underlying immune responses are further identified. Our results indicate that cross talks among these pathways and immune balances/competition between host and virus are always happened during the pathogenesis of PRRS. connected loop design was used to accommodate samples from 4 treatment groups.
Project description:Porcine reproductive and respiratory disease (PRRS) is the most important disease in swine industry worldwide. However, strategies such as vaccination and good biosecurity are not consistently successful to eliminate PRRSV. Though some interesting pathways have been tentatively examined recently, host molecular pathways utilized by PRRSV and the protective immune responses in resistant to PRRSV are largely unknown. In order to answer these questions, we herein characterize changes in global gene expressions in multiple tissues [tonsil, tracheobronchial lymph nodes (TBLN), Cranial lung (CR Lung), and distal lung (D Lung)] in response to PRRSV of high and low virulence. Both vaccinated and unvaccinated pigs are used for this study. Based on Ingenuity Pathway Analysis (IPA), molecule bases of some “black boxes” underlying immune responses are further identified. Our results indicate that cross talks among these pathways and immune balances/competition between host and virus are always happened during the pathogenesis of PRRS.
Project description:Porcine reproductive and respiratory syndrome virus (PRRSV) is a causative agent of an important infectious disease causing serious economic losses to swine industry called PRRS (porcine reproductive and respiratory syndrome). The clinical signs of this syndrome indcude respiratory disorders, abortions and variable mortality in piglets. To compare the virulence of highly diverse East European strains belonging to subtype 2 (Russian strain ILI and Belarusian strain BOR) and Danish strain from classical subtype 1 (DAN) the experimental study enrolling infection of piglets was performed. Gene expression profiles of peripheral blood mononuclear cells (PBMC) of piglets infected with three PRRSV strains vs control piglets were analysed by microarray analysis to gain insight into transcriptome changes after PRRSV infection.