Relative Contributions of Ebp Pili and the Collagen Adhesin Ace to Host Extracellular Matrix Protein Adherence and Experimental Urinary Tract Infection by Enterococcus faecalis OG1RF

Previous studies have demonstrated that the ebp operon and the ace gene of Enterococcus faecalis, encoding endocarditis- and biofilm-associated pili and an adhesin to collagen of E. faecalis, respectively, are both important in experimental urinary tract infections (UTI) and endocarditis. We have also shown that growth of E. faecalis in brain heart infusion (BHI) serum enhances Ebp pilus and Ace production and increases adherence to several host extracellular matrix proteins. Here, we report that deletion of ebpABC almost eliminated serum-elicited adherence to fibrinogen (P < 0.0001), resulted in moderate reduction in adherence to collagen (P < 0.05), and had no effect on fibronectin adherence relative to that of wild-type OG1RF. An OG1RF Delta ace Delta ebpABC double mutant showed further reduced collagen adherence versus that of the OG1RF Delta ace or OG1RF Delta ebpABC mutants (P < 0.001). These results were corroborated by complementation and/or studies with native pilus-enriched surface extracts and a collagen-secreting 3T6 fibroblast cell line, as well as antibody inhibition. In the UTI model, both the OG1RF Delta ace and OG1RF Delta ace Delta ebpABC mutants were found to be significantly attenuated compared to the wild type; however, no significant differences were observed between individual ace or ebp mutants and the OG1RF Delta ace Delta ebpABC mutant. In summary, these data implicate the Ebp pili as having some role in collagen adherence, albeit less than that of Ace, and a very major role in fibrinogen adherence, which may explain in part the importance of these pili in experimental endocarditis. The OG1RF Delta ace Delta ebpABC mutant was attenuated in the UTI model, although not significantly more so than the Delta ace or Delta ebpABC mutants, suggesting involvement of other E. faecalis factors in urinary tract colonization or infection.