Participating organizations (WP leader in bold): P7. IMR
Task 26.1 Production of VNN capsid protein (led by IMR). VNN outbreaks are mainly observed during early larval and juvenile stages of Atlantic halibut, when the immune system is not fully developed, and thus developing vaccines against this virus is especially challenging. We have several vaccine candidates: A vaccine with recombinant nodavirus capsid protein expressed in E.coli, which has previously been shown to elicit protection in turbot (Scophthalmus maximus), and to induce an immune response in Atlantic halibut with a protective character. Recombinant capsid protein has also been expressed in tobacco plants, from which it can be isolated as virus like particles (VLP) for integration in a vaccine. This is interesting as the plant (as a eukaryote), will presumably give post-translational modifications more in line with virus propagated in fish cells. We will work with two other eukaryotic expression systems, both suited for integration in an oral vaccine system. A formulation of the capsid protein of betanodavirus expressed recombinantly in E. coli has been shown to induce certain extent of protection in juvenile halibut. We will test the system for delivery of vaccine candidates through Artemia to Atlantic halibut larvae during late larval stages and the larvae will be challenged with VNN during early juvenile stages to assess for protection.
Task 26.2 Monitor and assess immune response and protection (led by IMR). We will monitor specific immune responses, viral load and possibly mortality at various time points following vaccination and further challenge with nodavirus. Samples will be taken after VNN exposure and examined by histology and immunohistochemistry for the presence of virus using specific antibodies against VNN. Since it is difficult to collect enough blood from juveniles in their early phase just after end metamorphosis, we will only concentrate on characterisation of specific cell mediated response by analysing a range cytokines associated with T cell responses (such as IL-12b, IFN-g, IL-1b) using qPCR assays for detection of virus and immune related genes that have already been established. Brain samples will be analysed for viral load.
Task 26.1 Production of VNN capsid protein (led by IMR). VNN outbreaks are mainly observed during early larval and juvenile stages of Atlantic halibut, when the immune system is not fully developed, and thus developing vaccines against this virus is especially challenging. We have several vaccine candidates: A vaccine with recombinant nodavirus capsid protein expressed in E.coli, which has previously been shown to elicit protection in turbot (Scophthalmus maximus), and to induce an immune response in Atlantic halibut with a protective character. Recombinant capsid protein has also been expressed in tobacco plants, from which it can be isolated as virus like particles (VLP) for integration in a vaccine. This is interesting as the plant (as a eukaryote), will presumably give post-translational modifications more in line with virus propagated in fish cells. We will work with two other eukaryotic expression systems, both suited for integration in an oral vaccine system. A formulation of the capsid protein of betanodavirus expressed recombinantly in E. coli has been shown to induce certain extent of protection in juvenile halibut. We will test the system for delivery of vaccine candidates through Artemia to Atlantic halibut larvae during late larval stages and the larvae will be challenged with VNN during early juvenile stages to assess for protection.
Task 26.2 Monitor and assess immune response and protection (led by IMR). We will monitor specific immune responses, viral load and possibly mortality at various time points following vaccination and further challenge with nodavirus. Samples will be taken after VNN exposure and examined by histology and immunohistochemistry for the presence of virus using specific antibodies against VNN. Since it is difficult to collect enough blood from juveniles in their early phase just after end metamorphosis, we will only concentrate on characterisation of specific cell mediated response by analysing a range cytokines associated with T cell responses (such as IL-12b, IFN-g, IL-1b) using qPCR assays for detection of virus and immune related genes that have already been established. Brain samples will be analysed for viral load.
