A multi-epitope plant-made chimeric protein (LTBentero) targeting common enteric pathogens is immunogenic in mice

Abstract

Key message A plant-based multiepitopic protein (LTBentero) containing epitopes from ETEC, S. typhimurium, and V. parahaemolyticus was produced in plants cells and triggered systemic and intestinal humoral responses in immunized mice. Abstract Around 200 million people suffer gastroenteritis daily and more than 2 million people die annually in developing countries due to such pathologies. Vaccination is an alternative to control this global health issue, however new low-cost vaccines are needed to ensure proper vaccine coverage. In this context, plants are attractive hosts for the synthesis and delivery of subunit vaccines. Therefore, in this study a plant-made multiepitopic protein named LTBentero containing epitopes from antigens of enterotoxigenic E. coli, S. typhimurium, and V. parahaemolyticus was produced and found immunogenic in mice. The LTBentero protein was expressed in tobacco plants at up to 5.29 µg g −1 fresh leaf tissue and was deemed immunogenic when administered to BALB/c mice either orally or subcutaneously. The plant-made LTBentero antigen induced specific IgG (systemic) and IgA (mucosal) responses against LTB, ST, and LptD epitopes. In conclusion, multiepitopic LTBentero was functionally produced in plant cells, being capable to trigger systemic and intestinal humoral responses and thus it constitutes a promising oral immunogen candidate in the fight against enteric diseases.

Molecular farming: Expanding the field of edible vaccines for sustainable fish aquaculture

“…84,185,186 This procedure is not necessarily limited to a production of a single antigen, but several studies have focused on the preparation of so-called multivalent vaccines combining antigens of various pathogens or many epitopes of one pathogen. 49,[187][188][189] Importantly, plant production systems generate active recombinant antigens with adequate post-translational modifications.…”

“…In addition to determining the localization of the recombinant antigen, its uptake can also be influenced by the incorporation of sequences encoding molecular adjuvants (CTBs, LTBs, flagellin, CPPs) and the creation of unique fusion proteins 84,185,186 . This procedure is not necessarily limited to a production of a single antigen, but several studies have focused on the preparation of so‐called multivalent vaccines combining antigens of various pathogens or many epitopes of one pathogen 49,187–189 …”

What Does Plant-Based Vaccine Technology Offer to the Fight against COVID-19?

“…Given that plant systems have been successfully adopted for the production of multiepitope proteins, the generation of such vaccines against COVID-19 is considered highly viable [88][89][90]. Besides expressing chimeric proteins, multiple antigen expression can be achieved by transplastomic technologies (operon-like expression) or at the nuclear level using the picornaviral 2A sequence that allows releasing individual antigens from a single coding gene [91].…”

Plant-based Vaccines: The Future of Preventive Healthcare?

“…Every day, 200 million people in the world experience health problems due to gastroenteritis. In developing countries, more than 2 million people die annually from such enteric diseases [23]. It has been reported that a multiepitopic protein from the antigens of enterotoxigenic E. coli, S. typhimurium and V. parahaemolyticus bacteria for use against these diseases is expressed in fresh leaf tissue as much as 5.29 μg g −1 in tobacco (Nicotiana tabacum) plants by A. tumefaciens-mediated transformation.…”

“…It has been reported that a multiepitopic protein from the antigens of enterotoxigenic E. coli, S. typhimurium and V. parahaemolyticus bacteria for use against these diseases is expressed in fresh leaf tissue as much as 5.29 μg g −1 in tobacco (Nicotiana tabacum) plants by A. tumefaciens-mediated transformation. Plant-made LTBentero antigen was found to be immunogenic when administered orally or subcutaneously to BALB/c mice, this antigen was also able to induce specific IgG (systemic) and IgA (mucosal) responses against LTB, ST, and LptD epitopes [23]. Moreover, by regulating the quality processes of transgenic plant-based vaccines, it has been ensured that the quality differences in the production steps are minimized.…”