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| Status |
Public on Dec 09, 2005 |
| Title |
Gene Expression Profiling Following DNA Vaccination of Rainbow Trout against Infectious Hematopoietic Necrosis Virus |
| Platform organisms |
Oncorhynchus mykiss; Salmo salar |
| Sample organism |
Oncorhynchus mykiss |
| Experiment type |
Expression profiling by array
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| Summary |
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
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| Overall design |
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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| Contributor(s) |
Purcell MK, Nichols KM, Park LK, Kurath G, Thorgaard GH, Wheeler P, Hansen JD, Herwig RP, Winton JR |
| Citation(s) |
16426680 |
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| Submission date |
Oct 27, 2005 |
| Last update date |
Mar 16, 2012 |
| Contact name |
Krista Nichols |
| E-mail(s) |
kmnichol@purdue.edu
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| Phone |
765-496-6848
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| Organization name |
Purdue University
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| Department |
Biological Sciences
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| Street address |
915 W State Street
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| City |
West Lafayette |
| State/province |
IN |
| ZIP/Postal code |
47992 |
| Country |
USA |
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| Platforms (1) |
| GPL2899 |
GRASP salmonid array, 16k v. 1.0, March 2004 |
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| Samples (10)
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| GSM76936 |
muscle tissue from a control fish, injected with PBS, replicate 1 |
| GSM77073 |
muscle tissue from a control fish, injected with PBS, replicate 2 |
| GSM77074 |
muscle tissue from a control fish, injected with PBS, replicate 3 |
| GSM77075 |
muscle tissue from a fish injected with vector, replicate 1 |
| GSM77076 |
muscle tissue from a fish injected with vector, replicate 2 |
| GSM77077 |
muscle tissue from a fish injected with vector, replicate 3 |
| GSM77078 |
muscle tissue from a fish injected with IHNV DNA Vaccine, replicate 1 |
| GSM77079 |
muscle tissue from a fish injected with IHNV DNA Vaccine, replicate 2 |
| GSM77080 |
muscle tissue from a fish injected with IHNV DNA Vaccine, replicate 3 |
| GSM77081 |
muscle tissue from a fish injected with IHNV DNA Vaccine, replicate 4 |
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| Relations |
| BioProject |
PRJNA93571 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE3522_RAW.tar |
36.6 Mb |
(http)(custom) |
TAR (of TXT) |
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