- Email: [email protected]
Transcriptional response of Atlantic salmon (Salmo salar) after primary versus secondary exposure to infectious salmon anemia virus (ISAV)
Abstracts / Fish & Shellfish Immunology 34 (2013) 1635–1691
coded for a putative protein with 522 amino acid residues. The amino acid sequence is very similar to that of other fish LAOs. Gene expression profiling revealed that LAO occurs ubiquitously in Atlantic cod. Upon experimental infection with Vibrio anguillarum, LAO gene expression in cod skin was found to increase 2-fold. Our data suggests that this molecule may have a role in mucosal defense of Atlantic cod against bacteria. * Corresponding author. E-mail address: [email protected] (V. Kiron)
O-158. Comparative genomics of innate anti-viral responses in fish A. Krasnov 1, *, S. Afanasyev 1, 2, I. Jensen 3, S.M. Jørgensen 1. 1
Nofima AS, PO Box 5010, NO-1430 Ås, Norway; Friedrich-Loeffler-Institut, Federal Research Institute for Animal Health, Institute of Immunology, 17493 Greifswald-Insel Riems, Germany; 3 Norwegian College of Fisheries Science, University of Tromsø, N-9037 Tromsø, Norway 2
Abstract Innate antiviral immunity has been extensively studied in teleost fish and many virus responsive genes (VRG) were identified and characterized. Advances in genomics bring research to a new level. A large volume of data was accumulated from gene expression profiling in Atlantic salmon with oligonucleotide microarrays (ONM); studies covered fish with IPN, ISA, CMS, PD and HSMI. To date, the list of salmon VRG contains 137 genes and several genes, including the most highly ranked receptor transporting protein, have not been previously reported for the role in antiviral defence. Atlantic cod is the first aquaculture species with a completely sequenced genome. A genome-wide ONM was used for analyses of gene expression in the brain during infection with nervous necrosis virus. A large fraction of the upregulated genes (546 features) were known or expected to have immune functions and many of these were homologs to salmon VRG. Comparative studies between teleost species provided new knowledge about the evolution of innate antiviral immunity, which showed a remarkably rapid sequence divergence in comparison with the entire proteome. VRG emerged both before and after separation of teleosts and tetrapods, and among genes found exclusively in fish species there were multiple trim, gig1 and gig2. Several VRG, including RNA helicase RIG-I and chemokine c-x-x10, are present in cyprinid and salmonid fish but were lost in the phylogenetically advanced orders. Apparently, part of genes has acquired different functional roles in higher vertebrates. Homologs of several fish VRG show neural specific expression in mammals, e.g. sacsin and opioid receptor. Atlantic cod is characterized by selective expansion of several medium-sized multigene families with ribose binding domains. VRG included members of three large gene families: trim and genes encoding pry-spry and nacht domains. The latter two with respectively 52 and 114 members in Atlantic cod have gone through expansions in different groups of fish. These proteins most likely have ligand binding properties and their propagation could be linked to the loss of MHC class II in the Atlantic cod genome. Transcriptomic studies increased knowledge of regulation and functions of VRG. These genes are characterized by a rapid induction and low tissue specificity, and their expression levels are related to the viral load but not to resistance against known pathogens. Most VRG showed highly correlated expression with ifna but many can be stimulated in an interferon-independent mode. In addition to viral infections, VRG are regulated under various conditions, such as parasite infestation and differentiation of red blood cells. * Corresponding author. E-mail address: Aleksei.Krasnov@nofima.no (A. Krasnov)
O-292. Profiles of Penaeus monodon GUT proteins in response to ‘oral WSSVvaccines’ A. Kulkarni 1, M.F. Brinchmann 1, J.H.M.W. Rombout 1,2, I.S.B. Singh 3, V. Kiron 1,*.
1659
1 Faculty of Biosciences and Aquaculture, University of Nordland, Bodø, Norway; 2 Wageningen University, Wageningen, The Netherlands; 3 National Centre for Aquatic Animal Health, Cochin University of Science and Technology, Kochi, India
Abstract White spot syndrome virus (WSSV) is a major shrimp pathogen and it has been reported that short-lived protection in shrimp, against this virus, can be provided using ‘vaccines’ containing WSSV envelope protein VP28 or formalin-inactivated WSSV. Nevertheless, the mechanisms of protection through these vaccines are not well described. Therefore, the interactions with the shrimp immune system need to be ascertained, in order to improve immunogenicity of the vaccines. In the present study, the aforementioned candidate vaccines were orally intubated in P. monodon and the expression profiles of proteins in the gut were determined by employing two-dimensional gel electrophoresis (2-DE). The same technique was also used on shrimps infected orally with live-WSSV to identify the differences in protein profiles. Around 90 proteins spots, with an altered expression (at least 1.5 fold), were subjected to mass spectrometry, and further analysed using bioinformatic databases. Based on the biological Gene Ontology terms, these proteins were classified under energy production, phenol oxidase activity, apoptosis, serine proteases, intracellular transport or nucleic acid synthesis. Additionally, spot intensities of some immune relevant proteins were compared with their corresponding mRNA levels (determined by quantitative real-time PCR). The results identified proteins that could be linked to the shrimp's immune response to vaccines. * Corresponding author. E-mail address: [email protected] (V. Kiron)
O-218. Transcriptional response of Atlantic salmon (Salmo salar) after primary versus secondary exposure to infectious salmon anemia virus (ISAV) M. Laflamme 1, *, F. LeBlanc 1, J.R. Arseneau 1, S. Leadbeater 2, B. Glebe 2, N. Gagné 1. 1 Department of Fisheries & Oceans Canada, Gulf Fisheries Centre, Moncton, NB, Canada; 2 Department of Fisheries and Oceans, Biological Station, St Andrews, NB, Canada
Abstract Following an infection with a specific pathogen, the acquired immune system in many organisms retains a specific memory of the infectious agent, and thus protects itself form subsequent infections. For example, Atlantic salmon that have survived an infection with a low-virulence infectious salmon anemia virus (ISAV) isolate are less susceptible to subsequent ISAV infections. A greater understanding of the mechanisms and immunological components involved in this acquired protection against ISAV is fundamental for the development of efficacious vaccines and treatments against this pathogen. To better understand the components involved in this phenomenon, we have used an Atlantic salmon DNA microarray to study the global gene expression responses of Atlantic salmon that had survived an infection with a low-virulence ISAV isolate, during the course of a secondary infection, 18 months later, with a highvirulence ISAV isolate. Our results show a clear reduction of ISAV viral loads in head-kidney of secondarily infected fish compared to primary infected fish. Further, we note a lower-expression of many antiviral innate immunity genes in the secondarily infected fish, suggesting that the acquired system was immediately activated. These results provide great insight into immunity components involved during primary and secondary ISAV infection. * Corresponding author. E-mail address: mark.lafl[email protected] (M. Laflamme)
coded for a putative protein with 522 amino acid residues. The amino acid sequence is very similar to that of other fish LAOs. Gene expression profiling revealed that LAO occurs ubiquitously in Atlantic cod. Upon experimental infection with Vibrio anguillarum, LAO gene expression in cod skin was found to increase 2-fold. Our data suggests that this molecule may have a role in mucosal defense of Atlantic cod against bacteria. * Corresponding author. E-mail address: [email protected] (V. Kiron)
O-158. Comparative genomics of innate anti-viral responses in fish A. Krasnov 1, *, S. Afanasyev 1, 2, I. Jensen 3, S.M. Jørgensen 1. 1
Nofima AS, PO Box 5010, NO-1430 Ås, Norway; Friedrich-Loeffler-Institut, Federal Research Institute for Animal Health, Institute of Immunology, 17493 Greifswald-Insel Riems, Germany; 3 Norwegian College of Fisheries Science, University of Tromsø, N-9037 Tromsø, Norway 2
Abstract Innate antiviral immunity has been extensively studied in teleost fish and many virus responsive genes (VRG) were identified and characterized. Advances in genomics bring research to a new level. A large volume of data was accumulated from gene expression profiling in Atlantic salmon with oligonucleotide microarrays (ONM); studies covered fish with IPN, ISA, CMS, PD and HSMI. To date, the list of salmon VRG contains 137 genes and several genes, including the most highly ranked receptor transporting protein, have not been previously reported for the role in antiviral defence. Atlantic cod is the first aquaculture species with a completely sequenced genome. A genome-wide ONM was used for analyses of gene expression in the brain during infection with nervous necrosis virus. A large fraction of the upregulated genes (546 features) were known or expected to have immune functions and many of these were homologs to salmon VRG. Comparative studies between teleost species provided new knowledge about the evolution of innate antiviral immunity, which showed a remarkably rapid sequence divergence in comparison with the entire proteome. VRG emerged both before and after separation of teleosts and tetrapods, and among genes found exclusively in fish species there were multiple trim, gig1 and gig2. Several VRG, including RNA helicase RIG-I and chemokine c-x-x10, are present in cyprinid and salmonid fish but were lost in the phylogenetically advanced orders. Apparently, part of genes has acquired different functional roles in higher vertebrates. Homologs of several fish VRG show neural specific expression in mammals, e.g. sacsin and opioid receptor. Atlantic cod is characterized by selective expansion of several medium-sized multigene families with ribose binding domains. VRG included members of three large gene families: trim and genes encoding pry-spry and nacht domains. The latter two with respectively 52 and 114 members in Atlantic cod have gone through expansions in different groups of fish. These proteins most likely have ligand binding properties and their propagation could be linked to the loss of MHC class II in the Atlantic cod genome. Transcriptomic studies increased knowledge of regulation and functions of VRG. These genes are characterized by a rapid induction and low tissue specificity, and their expression levels are related to the viral load but not to resistance against known pathogens. Most VRG showed highly correlated expression with ifna but many can be stimulated in an interferon-independent mode. In addition to viral infections, VRG are regulated under various conditions, such as parasite infestation and differentiation of red blood cells. * Corresponding author. E-mail address: Aleksei.Krasnov@nofima.no (A. Krasnov)
O-292. Profiles of Penaeus monodon GUT proteins in response to ‘oral WSSVvaccines’ A. Kulkarni 1, M.F. Brinchmann 1, J.H.M.W. Rombout 1,2, I.S.B. Singh 3, V. Kiron 1,*.
1659
1 Faculty of Biosciences and Aquaculture, University of Nordland, Bodø, Norway; 2 Wageningen University, Wageningen, The Netherlands; 3 National Centre for Aquatic Animal Health, Cochin University of Science and Technology, Kochi, India
Abstract White spot syndrome virus (WSSV) is a major shrimp pathogen and it has been reported that short-lived protection in shrimp, against this virus, can be provided using ‘vaccines’ containing WSSV envelope protein VP28 or formalin-inactivated WSSV. Nevertheless, the mechanisms of protection through these vaccines are not well described. Therefore, the interactions with the shrimp immune system need to be ascertained, in order to improve immunogenicity of the vaccines. In the present study, the aforementioned candidate vaccines were orally intubated in P. monodon and the expression profiles of proteins in the gut were determined by employing two-dimensional gel electrophoresis (2-DE). The same technique was also used on shrimps infected orally with live-WSSV to identify the differences in protein profiles. Around 90 proteins spots, with an altered expression (at least 1.5 fold), were subjected to mass spectrometry, and further analysed using bioinformatic databases. Based on the biological Gene Ontology terms, these proteins were classified under energy production, phenol oxidase activity, apoptosis, serine proteases, intracellular transport or nucleic acid synthesis. Additionally, spot intensities of some immune relevant proteins were compared with their corresponding mRNA levels (determined by quantitative real-time PCR). The results identified proteins that could be linked to the shrimp's immune response to vaccines. * Corresponding author. E-mail address: [email protected] (V. Kiron)
O-218. Transcriptional response of Atlantic salmon (Salmo salar) after primary versus secondary exposure to infectious salmon anemia virus (ISAV) M. Laflamme 1, *, F. LeBlanc 1, J.R. Arseneau 1, S. Leadbeater 2, B. Glebe 2, N. Gagné 1. 1 Department of Fisheries & Oceans Canada, Gulf Fisheries Centre, Moncton, NB, Canada; 2 Department of Fisheries and Oceans, Biological Station, St Andrews, NB, Canada
Abstract Following an infection with a specific pathogen, the acquired immune system in many organisms retains a specific memory of the infectious agent, and thus protects itself form subsequent infections. For example, Atlantic salmon that have survived an infection with a low-virulence infectious salmon anemia virus (ISAV) isolate are less susceptible to subsequent ISAV infections. A greater understanding of the mechanisms and immunological components involved in this acquired protection against ISAV is fundamental for the development of efficacious vaccines and treatments against this pathogen. To better understand the components involved in this phenomenon, we have used an Atlantic salmon DNA microarray to study the global gene expression responses of Atlantic salmon that had survived an infection with a low-virulence ISAV isolate, during the course of a secondary infection, 18 months later, with a highvirulence ISAV isolate. Our results show a clear reduction of ISAV viral loads in head-kidney of secondarily infected fish compared to primary infected fish. Further, we note a lower-expression of many antiviral innate immunity genes in the secondarily infected fish, suggesting that the acquired system was immediately activated. These results provide great insight into immunity components involved during primary and secondary ISAV infection. * Corresponding author. E-mail address: mark.lafl[email protected] (M. Laflamme)