Identification of linear epitopes on the flagellar proteins of Clostridioides difficile

Clostridioides difficile (C. difficile) is an opportunistic anaerobic bacterium that causes severe diseases of the digestive tract of humans and animals. One of the possible methods of preventing C. difficile infection is to develop a vaccine. The most promising candidates for vaccine antigens are the proteins involved in the adhesion phenomena. Among them, the FliC and FliD are considered to be suitable candidates. In this paper, the FliC and FliD protein polypeptide epitopes were mapped in silico and by using PEPSCAN procedure. We identified four promising epitopes: ¹¹⁷QRMRTLS¹²³, ²⁰⁵MSKAG²⁰⁹ of FliC and ²²⁶NKVAS²³⁰, ³⁰⁶TTKKPKD³¹² of FliD protein. We showed that ¹¹⁷QRMRTLS¹²³ sequence is not only located in TLR5-binding and activating region, as previously shown, but forms an epitope recognized by C. difficile-infected patients' antibodies. ²⁰⁵MSKAG²⁰⁹ is a C. difficile-unique, immunogenic sequence that forms an exposed epitope on the polymerized flagella structure which makes it a suitable vaccine antigen. ²²⁶NKVAS²³⁰ and ³⁰⁶TTKKPKD³¹² are well exposed and possess potential protective properties according to VaxiJen analysis. Our results open the possibility to use these epitopes as suitable anti-C. difficile vaccine antigens.