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Overexpression and oral immunogenicity of a dengue antigen transiently expressed in Nicotiana benthamiana

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Abstract

The feasibility of a safe and cost-effective oral dengue vaccine approach was investigated. Consensus domain III of the dengue virus (DENV) serotypes 1–4 envelope glycoprotein was fused with cholera toxin B subunit (CTB–cEDIII) and cloned into TMV-based 3′ pro-module for high level expression. The construct was co-infiltrated with TMV-based 5′ pro-module and Integrase module into Nicotiana benthamiana leaves using vacuum agroinfiltration. CTB–cEDIII fusion protein directed to endoplasmic reticulum by C-terminal SEKEDL sequence was expressed at up to 8.4% of total soluble protein at 5 days-post infiltration (dpi). After oral immunization of mice with the transiently expressed plant-derived CTB–cEDIII, high titers of serum IgG and sIgA against both cEDIII and CTB were observed, peaking at week four. Sera from immunized mice reacted with each of the four DENV serotype-specific envelope proteins domain III. These results show that (i) the TMV-based pro-vector system is suitable for high-level expression of antigen in plant cells and (ii) the CTB fusion with cEDIII could be exploited as an oral vaccine candidate for induction of both systemic and mucosal immune responses to DENV.

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Acknowledgements

This study was supported by NRF-2014K1B1A1073861 through the National Research Foundation (NRF) funded by the Korean Ministry of Science, ICT & Future Planning and by the Advanced Production Technology Development Program (312037-05), Ministry of Agriculture, Food and Rural Affairs (714001-07), Republic of Korea.

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  1. Department of Molecular Biology, Chonbuk National University, Jeonju, Republic of Korea

    Nguyen-Xuan Huy & Mi-Young Kim

  2. Biology Department, University of Education, Hue University, 34 Le Loi, Hue, Vietnam

    Nguyen-Xuan Huy

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  1. Nguyen-Xuan Huy
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M-Y Kim and N-X Huy designed the overall study and wrote the manuscript. M-Y. Kim generated the constructs and N-X Huy performed expression, the animal experiments and analyzed the data.

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Correspondence to Mi-Young Kim.

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Communicated by Ming-Tsair Chan.

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Huy, NX., Kim, MY. Overexpression and oral immunogenicity of a dengue antigen transiently expressed in Nicotiana benthamiana . Plant Cell Tiss Organ Cult 131, 567–577 (2017). https://doi.org/10.1007/s11240-017-1306-4

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