Project description:Oral immunization of trout against infectious pancreatic necrosis virus (IPNV) have been recently reported by using a DNA vector coding the viral capsid VP2 gene encapsulated in alginate microspheres. We report here an study of the transcripts in head kidney and pyloric ceca 7 days after oral vaccination of rainbow trout by using a newly designed rainbow trout 60-mer oligo microarray focused in their immune-related genes.
Project description:Infectious pancreatic necrosis virus (IPNV) is an aquatic virus that causes acute infection in freshwater and marine fish. The stage-specific expression of TNFα regulates Bad/Bid-mediated apoptosis and RIP1/ROS-mediated secondary necrosis in IPNV-infected fish cells. Using microRNA microarray and real-time quantitative PCR assays, the expression patterns of microRNA were characterized in different replication stages of IPNV or stimulation of LPS.
Project description:Oral immunization of trout against infectious pancreatic necrosis virus (IPNV) have been recently reported by using a DNA vector coding the viral capsid VP2 gene encapsulated in alginate microspheres. We report here an study of the transcripts in head kidney and pyloric ceca 7 days after oral vaccination of rainbow trout by using a newly designed rainbow trout 60-mer oligo microarray focused in their immune-related genes. Trout were obtained from 4 different farms, one group of trout was defined as belonging to one of the farms. Each of the trout 4 groups were divided into two subgroups of 6 trout for each subgroup. Subgroup I was orally vaccinated with 10 M-BM-5l of suspension of the vaccine microspheres each containing 10 M-BM-5g of pcDNA-VP2 diluted in 10M-BM-5l of PBS, while subgroup II received similar microspheres suspension but pcDNA. Head kidney and pyloric ceca were collected from each subgroup and RNA was extracted.
Project description:To investigate differentially expressed genes (DEGs) upon challenge with infectious salmon anemia virus (ISAV) or infectious pancreatic necrosis virus (IPNV) after IFNα treatment, in comparison to untreated cells. To examine the differentially expressed genes (DEGs) in knock-out (KO) cells with specific IFN receptors knockout, following IFNα treatment and infection with ISAV or IPNV. The expression profiles of these KO cells were compared with non-KO cells also treated with IFNα and infected with ISAV or IPNV.
Project description:Infectious pancreatic necrosis virus (IPNV) is an aquatic virus that causes acute infection in freshwater and marine fish. The stage-specific expression of TNFα regulates Bad/Bid-mediated apoptosis and RIP1/ROS-mediated secondary necrosis in IPNV-infected fish cells. Using microRNA microarray and real-time quantitative PCR assays, the expression patterns of microRNA were characterized in different replication stages of IPNV or stimulation of LPS. Two-condition experiment, normal vs IPNV-infected cells (at 6, 12 or 24 hr post-infection), or normal vs LPS-stimulated cells (at 6, 12 or 24 h post-treatment).
Project description:Infectious pancreatic necrosis (IPN) is a serious viral disease that causes significant economic losses in salmon aquaculture. To characterize the host-pathogen relationship in IPN, we analysed transcriptional profiles of salmon head kidney (SHK-1) cells infected with infectious pancreatic necrosis virus (IPNV) at three timepoints over six days (at 1, 3 & 6 days post infection. The transcriptome was investigated using the TRAITS / SGP 16950-feature Atlantic salmon cDNA microarray, which is enriched for genes with functions related to the immune response.
Project description:Atlantic salmon juveniles were screened for swimming performance and separated into either poor or good swimmers. After ten weeks of rearing in fresh water, during which both swimming performance groups were part of an exercise training experiment, fish were transferred to seawater and challenged with infectious pancreatic necrosis virus (IPNV) in a co-habitation test. When mortality curve levelled out (45 days post commencement of challenge test), fish that had previously been categorized as good swimmers displayed a significantly higher survival (86.1%) compared to poor swimmers (77.6%). Global gene expression analyses were performed to search for disease resistance correlates. Cardiac ventricle expression of 21 genes was greater in poor swimmers than in good swimmers. These genes were previously classified as virus-responsive genes (VRGs), being reliable markers of viral load. This suggested that inherent swimming performance is associated with higher disease resistance. Atlantic salmon post-smolts belonging to groups previously classified as either poor or good swimmers were challenged with infectious pancreatic necrosis virus (IPNV). Heart ventricle was sampled from challenged and unchallenged fish on day 45 post-commencement of the challenge (when no more mortalities were registered). Nine poor swimmers and nine good swimmers were hybridized against a common reference sample composed of nine unchallenged fish.
Project description:Infectious pancreatic necrosis virus (IPNV) is a fish-derived pathogen and possess a bi-segmented, double-stranded RNA genome. By using zebrafish 14K oligo-microarray and quantitative RT-PCR, we identified differential expression of a defined subset of genes involved in apoptosis and immunity at 6-, 12- and 24- hour after IPNV infection. Transcripts divided into 11 functional categories were significantly modulated by IPNV, including immune response, apoptosis, transcription, signal transduction, lipid and cholesterol metabolism, carbohydrate metabolism, oxidative phosphorylation, cell cycle, protein degradation, protein folding and stress response, protein synthesis, nucleoside metabolism and synthesis. Most of pro-apoptotic bcl-2 family members were up-regulated after IPNV infection. Activation of pro-apoptotic members might disrupt potential of mitochondria and leaded to the mitochondria-mediated apoptosis in the late stage of IPNV infection. After treating the IPNV-infected cells with TNFα inhibitor AP126, expression of two bcl-2 family genes Bad and Bid and activation of caspase-8 and -3 had been inhibited significantly in early stage of IPNV infection. Expression of RIP-1 and Bmf-1 these two necroptosis-related genes and production of ROS were diminished in virus-infected cells which pre-treated with AP126. Our study shows the interactions between host cells and IPNV, the molecular mechanisms involved in IPNV-induced pathogenesis, and the variation of transcriptome through TNFα during infection which shows the important component of host defense. TNFα might lead to apoptosis in early stage and necrosis in late stage in ZF4 cells infected by IPNV. TNFα is crucial to apoptosis and ROS-mediated necrosis caused by IPNV in zebrafish cells.
