Immunogenicity of a lettuce-derived vaccine candidate expressing the E2 protein against classical swine fever virus
2013
Classical swine fever (CSF) is a highly infectious hemorrhagic disease that affects domestic and wild swine. Live attenuated vaccines for classical swine fever viruses (CSFV) can provide rapid onset protection against CSF. However, because of serious safety concerns regarding the use of modified-live virus vaccines, the development of novel CSFV vaccines is urgently required. The E2 protein plays a vital role in viral replication and is essential in CSF development; thus represents an ideal vaccine target. Transgenic plants provide cost-effective and scalable systems for the production and delivery of viral protein, and might have potential use as oral vaccines. In this study, we generated a transgenic lettuce expressing the CSFV E2 gene, under the control of the CaMV 35S promoter, using Agrobacterium-mediated transformation. We confirmed the stable integration and transcriptional expression of the E2 gene using genomic DNA PCR amplification and quantitative real-time PCR analysis, respectively. We then confirmed E2 expression using Western blot analysis. Based on ELISA analysis, levels of E2 protein in leaf tissues of different transgenic lines ranged between 0.21 and 0.53 μg mg⁻¹ of the total soluble protein. We detected E2-specific antibodies in mice following their subcutaneous and oral immunization with protein extracts from transgenic lettuce. Following the vaccination of mice with the recombinant E2 protein, we identified upregulated IFN-γ and IL-10 in their splenocytes.
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