Unknown,Transcriptomics,Genomics,Proteomics

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Infectious hematopoietic necrosis virus (IHNV) persistence in Sockeye Salmon: effect on brain transcriptome and response to the viral mimic poly(I:C)

ABSTRACT: To characterise the transcriptional response in brain of Sockeye Salmon (Oncorhynchus nerka) that were persistently infected with infectious hematopoietic necrosis virus (IHNV) and to determine whether carrying the IHNV affects the ability to respond to other immunological challenges we compared the brain transcriptome of IHNV carriers, IHNV-negative survivors, and naïve Sockeye Salmon that were never exposed to IHNV. In addition we determined the transcriptional changes among carriers, survivors and naïve fish in their response to the viral mimic polyriboinosinic polyribocytidylic acid (poly(I:C)), using the cGRASP 44K salmon oligoarray. Fish that survived an immersion challenge with IHNV and time-matched control (naïve) fish that were never exposed to IHNV were reassigned into 2 tanks each at 271 days post challenge (dpc). One group each of the challenged and naïve fish received an intraperitoneal injection of poly(I:C). For controls, a group each of challenged and naïve fish were not injected. Brain tissues were sampled from each tank at 3 (274 dpc), 7 (278 dpc) and 10 (281 dpc) days after poly(I:C) injection. Based on the presence of IHNV in brain fish were assigned into “carriers” and “survivors”, i.e. fish with detectable and undetectable levels of IHNV, respectively. For transcriptomic profiling brain samples were analysed from naïve (n = 7), survivors (n = 7), and carriers (n = 5), as well as poly(I:C)-injected naïve (n = 7), poly(I:C)-injected survivors (n = 7), and poly(I:C)-injected carriers (n = 2). All analysed brain samples were collected 274 dpc (i.e. 3 d after poly(I:C) injection) with the exception of two carriers that were collected 278 dpc. The smaller sample size for carriers was a result of the few individuals that survived the exposure that ended up carrying the virus.

ORGANISM(S): Oncorhynchus nerka

SUBMITTER: Ben Koop

PROVIDER: E-GEOD-65241 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:The Infectious Hematopoietic Necrosic Virus (IHNV) DNA vaccine is based on the viral glycoprotein gene (G gene) and induces a non-specific anti-viral immune response and long-term specific immunity against IHNV in salmonid fishes. This study characterized gene expression responses associated with the early anti-viral response. Homozygous rainbow trout were injected intra-muscularly (I.M.) with vector DNA or the IHNV DNA vaccine. Gene expression at the I.M. site was evaluated using a 16,000 feature salmon cDNA microarray. Eighty different transcripts were significantly modulated in the vector DNA group while 910 transcripts were modulated in the IHNV DNA vaccinated group relative to control group. Quantitative reverse transcriptase PCR was used to examine expression of selected genes at the I.M. site and in other secondary tissues. In the localized response (I.M. site), the magnitudes of gene expression changes were higher in the vaccinate group relative to the vector DNA group for the majority of genes analyzed. At secondary systemic sites (e.g. gill, kidney and spleen) evaluated with RT-PCR of specific genes, the main difference was the up-regulation of the type I IFN-related genes in only the IHNV DNA vaccinated group. Keywords: disease vaccine response Gene expression at the site of I.M injection was evaluated 7 days post-injection in clonal rainbow trout. Gene expression was compared across three treatments, with replicate clonal rainbow trout individuals (Hot Creek strain) within each treatment. Treatments included injection with 1X PBS (control group), plasmid vector only (without IHNV G gene; hereafter referred to as vector group), and the pIHNV vaccine (vaccine group). Three clonal individuals from the control, three individuals from the vector group, and four individuals from the vaccine group were labeled with Cy3 and used for individual hybridizations to ten slides, comparing hybridization of each individual to a major reference RNA pool. The major reference pool consisted of three unhandled clonal rainbow trout and constituted a pool of RNA from the anterior kidney, spleen, liver and muscle and was labelled with Cy5. Total RNA was extracted from samples, amplified, and hybridized to 16,000 feature Atlantic salmon cDNA arrays developed by the Genomic Research for Atlantic Salmon project. Differential expression was compared among control, vector, and vaccine groups.

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Infectious hematopoietic necrosis virus (IHNV) persistence in Sockeye Salmon: effect on brain transcriptome and response to the viral mimic poly(I:C)

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