Sammendrag

The plant pathogenic fungus Fusarium langsethiae produces the highly potent mycotoxins HT-2 and T-2. Since these toxins are frequently detected at high levels in oat grain lots, they pose a considerable risk for food and feed safety in Norway, as well as in other north European countries. To reduce the risk of HT-2/T- 2-contaminated grain lots to enter the food and feed chain, it is important to identify factors that influence F. langsethiae infection and mycotoxin development in oats. However, the epidemiology of F. langsethiae is unclear. A three-year survey was performed to reveal more of the life cycle of F. langsethiae and its interactions with oats, other Fusarium species, as well as insects, mites and weeds. We searched for inoculum sources by quantifying the amount of F. langsethiae DNA in crop residues, weeds, and soil sampled from a selection of oat-fields. To be able to define the onset of infection, we analysed the amount of F. langsethiae DNA in oat plant material sampled at selected growth stages (between booting and maturation), as well as the amount of F. langsethiae DNA and HT-2 and T-2 toxins in the mature grain. We also studied the presence of possible insect- and mite vectors sampled at the selected growth stages using Berlese funnel traps. The different types of materials were also analysed for the presence F. graminearum DNA, the most important deoxynivalenol producer observed in Norwegian cereals, and which presence has shown a striking lack of correlation with the presence of F. langsethiae in oat. Results show that F. langsethiae DNA may occur in the oat plant already before heading and flowering. Some F. langsethiae DNA was observed in crop residues and weeds, though at relatively low levels. No Fusarium DNA was detected in soil samples. Of the arthropods that were associated with the collected oat plants, aphids and thrips species were dominating. Further details will be given at the meeting.

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Chronic Hepatitis B Virus (HBV) infection leads to severe liver pathogenesis associated with significant morbidity and mortality. As no curable medication is yet available, vaccination remains the most costeffective approach to limit HBV spreading and control the infection. Although safe and efficient, the standard vaccine based on production of the small (S) envelope protein in yeast fails to elicit an effective immune response in about 10% of vaccinated individuals, which are at risk of infection. One strategy to address this issue is the development of more immunogenic antigens. Here we describe a novel HBV antigen obtained by combining relevant immunogenic determinants of S and large (L) envelope proteins. Our approach was based on the insertion of residues 21-47 of the preS1 domain of the L protein (nomenclature according to genotype D), involved in virus attachment to hepatocytes, within the external antigenic loop of S. The resulting S/preS121-47 chimera was successfully produced in HEK293T and Nicotiana benthamiana plants, as a more economical recombinant protein production platform. Comparative biochemical, functional and electron microscopy analysis indicated assembly of the novel antigen into subviral particles in mammalian and plant cells. Importantly, these particles preserve both S- and preS1-specific epitopes and elicit significantly stronger humoral and cellular immune responses than the S protein, in both expression systems used. Our data promote this antigen as a promising vaccine candidate to overcome poor responsiveness to the conventional, S protein-based, HBV vaccine.

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The hepatitis C virus (HCV) is a major etiologic agent for severe liver diseases ( e.g . cirrhosis, fibrosis and hepatocellular carcinoma). Approximately 140 million people have chronic HCV infections and about 500 000 die yearly from HCV-related liver pathologies. To date, there is no licensed vaccine available to prevent HCV infection and production of a HCV vaccine remains a major challenge. Here, we report the successful production of the HCV E1E2 heterodimer, an important vaccine candidate, in an edible crop (lettuce, Lactuca s ativa ) using Agrobacterium - mediated transient expression technology. The wild-type dimer (E1E2) and a variant without an N-glycosylation site in the E2 polypeptide (E1E2 Δ N6) were expressed, and appropriate N-glycosylation pattern and functionality of the E1E2 dimers were demonstrated. The humoral immune response induced by the HCV proteins was investigated in mice following oral administration of lettuce antigens with or without previous intramuscular prime with the mammalian HEK293T cell-expressed HCV dimer. Immunization by oral feeding only resulted in development of weak serum levels of anti-HCV IgM for both antigens; however, the E1E2 Δ N6 proteins produced higher amounts of secretory IgA, suggesting improved immunogenic properties of the N-glycosylation mutant. The mice group receiving the intramuscular injection followed by two oral boosts with the lettuce E1E2 dimer developed a systemic but also a mucosal immune response, as demonstrated by the presence of anti-HCV secretory IgA in faeces extracts. In summary, our study demonstrates the feasibility of producing complex viral antigens in lettuce, using plant transient expression technology, with great potential for future low-cost oral vaccine development.

Sammendrag

The plant pathogenic fungus Fusarium langsethiae produces the highly potent mycotoxins HT-2 and T-2. Since these toxins are frequently detected at high levels in oat grain lots, they pose a considerable risk for food and feed safety in Norway, as well as in other north European countries. To reduce the risk of HT-2/T- 2-contaminated grain lots to enter the food and feed chain, it is important to identify factors that influence F. langsethiae infection and mycotoxin development in oats. However, the epidemiology of F. langsethiae is unclear. A three-year survey was performed to reveal more of the life cycle of F. langsethiae and its interactions with oats, other Fusarium species, as well as insects, mites and weeds. We searched for inoculum sources by quantifying the amount of F. langsethiae DNA in weeds, crop residues, and soil, sampled from a predetermined selection of oat-fields. To be able to define the onset of infection, we analysed the amount of F. langsethiae DNA in oat plant material sampled at selected growth stages (between booting and maturation), as well as the amount of F. langsethiae DNA and HT-2 and T-2 toxins in the mature grain. We also studied the presence of possible insect- and mite vectors sampled at the selected growth stages using Berlese funnel traps. All the different types of materials were also analysed for the presence F. graminearum DNA, the most important deoxynivalenol producer observed in Norwegian cereals, and which presence has shown a striking lack of correlation with the presence F. langsethiae in oat. Preliminary results show that F. langsethiae DNA may occur in the oat plant before heading and flowering. Some F. langsethiae DNA was observed in crop residues and weeds, though at relatively low levels. More results from this work will be presented at the meeting.

Sammendrag

The plant pathogenic fungus Fusarium langsethiae produces the highly potent mycotoxins HT-2 and T-2. Since these toxins are frequently detected at high levels in oat grain lots, they pose a considerable risk for food and feed safety in Norway, as well as in other north European countries. To reduce the risk of HT-2/T- 2-contaminated grain lots to enter the food and feed chain, it is important to identify factors that influence F. langsethiae infection and mycotoxin development in oats. However, the epidemiology of F. langsethiae is unclear. A three-year survey was performed to reveal more of the life cycle of F. langsethiae and its interactions with oats, other Fusarium species, as well as insects, mites and weeds. We searched for inoculum sources by quantifying the amount of F. langsethiae DNA in weeds, crop residues, and soil, sampled from a predetermined selection of oat-fields. To be able to define the onset of infection, we analysed the amount of F. langsethiae DNA in oat plant material sampled at selected growth stages (between booting and maturation), as well as the amount of F. langsethiae DNA and HT-2 and T-2 toxins in the mature grain. We also studied the presence of possible insect- and mite vectors sampled at the selected growth stages using Berlese funnel traps. All the different types of materials were also analysed for the presence F. graminearum DNA, the most important deoxynivalenol producer observed in Norwegian cereals, and which presence has shown a striking lack of correlation with the presence F. langsethiae in oat. Preliminary results show that F. langsethiae DNA may occur in the oat plant before heading and flowering. Some F. langsethiae DNA was observed in crop residues and weeds, though at relatively low levels. More results from this work will be presented at the meeting.

Sammendrag

The global spread of dengue fever threatens a large percentage of the world’s population. The disease causes great human suffering, a high mortality from dengue haemorrhagic fever and its complications, and major costs. There is currently no vaccine to prevent dengue virus infection. Our project aims to express a tetravalent vaccine candidate in tobacco chloroplasts, a cost effective system, and hence to contribute to innovation and bio-economy as a long term goal.