Lactobacilli play an important role in the maintenance of a healthy vaginal microbiota， and some select species are widely used as probiotics. Vaginal isolates of DSM 14869 and DSM 14870 were previously selected to develop the probiotic EcoVag capsules and showed therapeutic effects in women with bacterial vaginosis (BV). However， the molecular mechanisms involved in their probiotic activity are largely unknown. In this study， we identified three cell surface molecules in DSM 14869 that promote adhesion to vaginal epithelial cells (VEC) by constructing dedicated knockout mutants， including exopolysaccharides (EPSs)， a protein containing MucBP-like domains (N506_1778)， and a putative novel adhesin... More
Lactobacilli play an important role in the maintenance of a healthy vaginal microbiota， and some select species are widely used as probiotics. Vaginal isolates of DSM 14869 and DSM 14870 were previously selected to develop the probiotic EcoVag capsules and showed therapeutic effects in women with bacterial vaginosis (BV). However， the molecular mechanisms involved in their probiotic activity are largely unknown. In this study， we identified three cell surface molecules in DSM 14869 that promote adhesion to vaginal epithelial cells (VEC) by constructing dedicated knockout mutants， including exopolysaccharides (EPSs)， a protein containing MucBP-like domains (N506_1778)， and a putative novel adhesin (N506_1709) with rib/alpha-like domain repeats. EPS knockout mutants revealed 20-fold and 14-fold increases in adhesion to Caco-2 and HeLa cells， respectively， compared with wild type， while the adhesion to VEC was reduced 30% by the mutation， suggesting that EPSs might mediate tissue tropism for vaginal cells. A significant decrease in adhesion to Caco-2 cells， HeLa cells， and VEC was observed in the N506_1778 knockout mutant. The N506_1709 mutant showed no significant difference for the adhesion to Caco-2 and HeLa cells compared with wild type (WT); in contrast， the adhesion to VEC revealed a significant decrease (42%)， suggesting that N506_1709 might mediate specific binding to stratified squamous epithelial cells， and this putative novel adhesin was annotated as vaginal epithelium adhesin (LVEA). Thus， we have discovered an important role for EPSs and a novel adhesin， LVEA， in the adhesive capacity of a vaginal probiotic strain. Lactobacilli are known to contribute to the maintenance of a healthy vaginal microbiota and some are selected as probiotics for the prevention or treatment of urogenital diseases， such as bacterial vaginosis. However， the molecular mechanisms for these health-promoting effects are not fully understood. Here， we functionally identified three cell surface factors of a strain potentially involved in its adhesion to vaginal epithelial cells， including exopolysaccharides (EPSs) and two sortase-dependent proteins (N506_1778 and N506_1709). We could demonstrate the tissue-specific adhesion of EPSs to vaginal cells and that N506_1709 might be a novel adhesin specifically mediating bacterial binding to stratified squamous epithelial cells. The results provide important new information on the molecular mechanisms of vaginal spp. adhesion.
