Biotechnology and Bioprocess Engineering 2019; 24(6): 954-963  
Immunogenicity of Fusion Protein of Cholera Toxin B Subunit-Porphyromonas gingivalis 53-kDa Minor Fimbrial Protein Produced in Nicotiana benthamiana
Tae-Geum Kim1, Tran Thuy Lan2,3, and Jin-Yong Lee3,*
1Center for Jeongup Industry-Academy-Institute Cooperation, Chonbuk National University, Jeongup 56212, Korea
2Department of Molecular Biology, Chonbuk National University, Jeonju 54896, Korea
3Institute of Biotechnology, Hue University, Hue, Vietnam
4Department of Oral Microbiology, School of Dentistry, Kyung Hee University, Seoul 02447, Korea
Correspondence to: Jin-Yong Lee*
Department of Oral Microbiology, School of Dentistry, Kyung Hee University, Seoul 02447, Korea
Tel: +82-2-960-2838; Fax: +82-2-960-2838
E-mail: ljinyong@khu.ac.kr
Received: May 11, 2019; Revised: July 29, 2019; Accepted: July 31, 2019; Published online: December 31, 2019.
© The Korean Society for Biotechnology and Bioengineering. All rights reserved.

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
Abstract
Porphyromonas gingivalis induces destruction of periodontal tissues that surround and support the teeth, contributing to the development of periodontitis, which eventually results in tooth loss in adults. A 53-kDa protein of P. gingivalis is a major subunit variant protein of minor fimbriae (Mfa1), and is reported to be highly immunogenic and considered as a potential vaccine candidate. The gene encoding 53-kDa protein, was divided into three segments, and each DNA segment was fused to the gene coding for cholera toxin B subunit (CTB) to improve gut mucosal immune responses. The ctb-mfa1 fusion genes were expressed in the leaf tissues of Nicotiana benthamiana using agroinfiltration. Immunoblot analysis revealed that CTB-Mfa1 fusion proteins were produced in the agroinfiltrated leaves. The interaction of the plant-produced CTB-Mfa1 fusion proteins with GM1-ganglioside, which acts as the binding site for native CTB, was confirmed by GM1-ELISA. Mice immunized orally with the agroinfiltrated leaf powder containing the CTB-Mfa1 fusion proteins elicited serum IgG and fecal IgA antibodies to CTB and Mfa1. These results suggest that CTB-Mfa1 fusion proteins produced in plants can be used as an oral vaccine to control P. gingivalis infection-associated periodontitis.
Keywords: Porphyromonas gingivalis, 53-kDa minor fimbrial protein, periodontitis, agroinfiltration, plant-based vaccine


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