Flagellin-adjuvanted trivalent mucosal vaccine targeting key periodontopathic bacteria

Periodontal disease (PD) is caused by microbial dysbiosis and accompanying adverse inflammatory responses. Due to its high incidence and association with various systemic diseases, disease-modifying treatments that modulate dysbiosis serve promising therapeutic approaches. In this study, to simulate the pathophysiological situation, we established a ‘temporary ligature plus oral infection model’ that incorporates temporary silk ligature and oral infection with the cocktail of live Tannerella forsythia ( Tf ), Pophyromonas gingivalis ( Pg ), and Fusobacterium nucleatum ( Fn ) in mice and tested the efficacy of a new trivalent mucosal vaccine. It has been reported that Tf , a red complex pathogen, amplifies periodontitis severity by interacting with periodontopathic bacteria such as Pg and Fn. Here we developed a recombinant mucosal vaccine targeting a surface-associated protein BspA of Tf by genetically combining truncated BspA with built-in adjuvant flagellin (FlaB). To simultaneously induce Tf -, Pg -, and Fn -specific immune responses, it was formulated as a trivalent mucosal vaccine containing Tf -FlaB-tBspA (BtB), Pg -Hgp44-FlaB (HB), and Fn -FlaB-tFomA (BtA). Intranasal immunization with the trivalent mucosal vaccine (BtB+HB+BtA) prevented alveolar bone loss and gingival pro-inflammatory cytokine production. Vaccinated mice exhibited significant induction of Tf- tBspA-, Pg- Hgp44-, and Fn- tFomA-specific IgG and IgA responses in the serum and saliva, respectively. The anti-sera and anti-saliva efficiently inhibited epithelial cell invasion by Tf and Pg and interfered with biofilm formation by Fn . In summary, the flagellin-adjuvanted trivalent mucosal vaccine-mediated immunomodulation would serve as a promising choice for clinically managing dysbiotic bacteria-induced periodontitis.