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Transgenic plants expressing a Clostridium difficile spore antigen as an approach to develop low‐cost oral vaccines

Gastrointestinal infections caused by Clostridium difficile lead to significant impact in terms of morbidity and mortality, causing from mild symptoms, such as a low‐grade fever, watery stools, and minor abdominal cramping as well as more severe symptoms such as bloody diarrhea, pseudomembrane colitis, and toxic megacolon. Vaccination is a viable approach to fight against C. difficile and several efforts in this direction are ongoing. Plants are promising vaccine biofactories offering low cost, enhanced safety, and allow for the formulation of oral vaccines. Herein, the CdeM protein, which is a spore antigen associated with immunoprotection against C. difficile, was selected to begin the development of plant‐based vaccine candidates. The vaccine antigen is based in a fusion protein (LTB‐CdeM), carrying the CdeM antigen, fused to the carboxi‐terminus of the B subunit of the Escherichia coli heat‐labile enterotoxin (LTB) as a mucosal immunogenic carrier. LTB‐CdeM was produced in plants using a synthetic optimized gene according codon usage and mRNA stability criteria. The obtained transformed tobacco lines produced the LTB‐CdeM antigen in the range of 52–90 μg/g dry weight leaf tissues. The antigenicity of the plant‐made LTB‐CdeM antigen was evidenced by GM1‐ELISA and immunogenicity assessment performed in test mice revealed that the LTB‐CdeM antigen is orally immunogenic inducing humoral responses against CdeM epitopes. This report constitutes the first step in the development of plant‐based vaccines against C. difficile infection.