Abstract
Dengue virus (DENV) infection is an emerging global health threat. DENV consists of four distinct serotypes, necessitating a tetravalent vaccine. In this study, expression of consensus envelope protein domain III (cEDIII) fused to cholera toxin B subunit (CTB) in transgenic rice calli was improved using the luminal binding protein BiP at the N-terminus and the SEKDEL signal sequences at the C-terminus, targeting the recombinant protein to endoplasmic reticulum (ER). We found that the fusion protein showed higher levels of expression when compared to the fusion proteins using rice amylase 3D (RAmy3D) or CTB native signal sequence only. The CTB-cEDIII fusion protein was evaluated as an oral dengue vaccine candidate in mice. Serotype specific systemic IgG antibodies and specific IgA response in feces were detected and furthermore, T cell proliferation and high frequency antibody-secreting B cells were detected in the spleen. These results suggest the possible use of plant-based dengue tetravalent vaccine targeted to the mucosal immune system for induction of systemic and mucosal immune responses to DENV infection.
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Acknowledgments
This study was supported by NRF-2014K1B1A1073861 through the National Research Foundation (NRF) funded by Korean Ministry of Science, ICT & Future Planning. The study was also supported by the BK21 Plus program in the Departments of Bioactive Material Sciences award to Mr. B-Y Kim. We thank Dr Hwang (POSTECH, Pohwang, Korea) for providing the genomic DNA of BiP signal sequence, Dr. Kim (Chonbuk National University) for scientific advice and Mr. Park (Chonbuk National University) for helping with the analysis of cellular responses.
Author contributions
M-Y. Kim, M-S. Yang and Y-S. Jang conceived and designed the overall study. M-Y. Kim and S-M Oh generated the constructs and performed expression experiments. B-Y. Kim performed the animal experiments and analysed the data. M-S. Yang, Y-S. Jang and R.R. advised on dengue vaccine strategy. M-Y. Kim, B-Y. Kim and R. R. wrote the manuscript.
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Mi-Young Kim and Beyng-Young Kim have contributed equally to this study.
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Kim, MY., Kim, BY., Oh, SM. et al. Oral immunisation of mice with transgenic rice calli expressing cholera toxin B subunit fused to consensus dengue cEDIII antigen induces antibodies to all four dengue serotypes. Plant Mol Biol 92, 347–356 (2016). https://doi.org/10.1007/s11103-016-0517-0
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DOI: https://doi.org/10.1007/s11103-016-0517-0