Heterogeneous expression of Pil3 pilus is critical for Streptococcus gallolyticus translocation across polarized colonic epithelial monolayers

Streptococcus gallolyticus is an opportunistic pathogen responsible for septicemia and endocarditis. We report that S. gallolyticus UCN34 adheres and crosses epithelial monolayers in a Pil3 dependent manner. Confocal images revealed a paracellular passage. Both the Δpil3 mutant and the Pil3+ overexpressing variant were unable to cross Caco-2 and T84 barriers. However, combining live Δpil3 mutant with fixed Pil3+ variant in a 9:1 ratio allowed efficient translocation of the Δpil3 mutant. These results demonstrate that heterogeneous expression of Pil3 plays a key role for UCN34 translocation across the intestinal barrier. Through this skilful strategy, S. gallolyticus probably evade host immune responses.