Plant expression of antigens of animal pathogens and its application for oral vaccination

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This project was aiming at expression of genes from animal pathogenic microbes, and development of mucosal vaccine as an injectable form into nasal or oral cavities in combination with cytokines. As model antigens, salivary antigens of ticks that transmit various protozoan parasite were investigated. Those antigens combined with cytokines such as interleukin-1 or -2 will be used as tick waccines.We introduced a bovine transforming growth factor beta gene into tobacco by using Agrobacterium system, and confirmed its expression in regenerated plant. Expression was enhanced by addition of legumine signal sequence. We also expressed bovine IL-1 and IL-2 in tobacco plants. Biological activities of these cytokines were confirmed by in vitro assays.Genes for tick cement proteins, serine proteinases, and their inhibitors (serpin) were cloned and their molecular structures were investigated. Biological activities of these molecules were also demonstrated. Vaccination of rabbit with recombinant cement proteins inhibited tick infestation, which suggested these proteins are good candidate for commercial production of tick vaccines.