Mechanisms of decreased susceptibility to beta-defensins by Treponema denticola.
Treponema denticola, a periodontal pathogen, is relatively resistant to human beta-defensins, which are small cationic antimicrobial peptides produced by a number of cells, including the gingival epithelium. Using two independent methods, we previously demonstrated that T. denticola proteases are not responsible for decreased vulnerability to defensins. In this study, we confirmed that the major outer membrane protease, dentilisin, is not responsible for T. denticola insensitivity to defensins and examined several other possible mechanisms, including reduced binding to the bacterial surface and efflux pump activity. It has been suggested that some bacteria mask their surfaces with serum proteins. T. denticola grown in a serum-free medium did not exhibit increased susceptibility to human beta-defensin 2 and 3 (hbetaD-2 and hbetaD-3, respectively), suggesting that cloaking of the outer surface with host proteins is not involved in defensin resistance. Nonetheless, we demonstrated that T. denticola binds significantly less hbetaD-2 and -3 than susceptible organisms bind, suggesting that the unusual outer membrane composition of T. denticola may discourage cationic peptide binding. Efflux pumps have been shown to mediate resistance to antibiotics and cationic peptides in other bacteria, and their role in T. denticola's relative resistance to beta-defensins was investigated. Three inhibitors of bacterial ATP-binding cassette (ABC) efflux pumps had no effect on T. denticola's susceptibility to hbetaD-2 or -3. In contrast, a proton motive force inhibitor, carbonyl cyanide 3-chlorophenylhydrazone, increased the susceptibility of T. denticola to killing by hbetaD-3, demonstrating a potential role for efflux pumps (other than ABC pumps) in resistance to this peptide. Our data suggest that the combination of decreased defensin binding and efflux of any peptide which enters the cytoplasm may explain T. denticola's relative resistance to human beta-defensins.
Brissette
Catherine A
CA
Department of Pathobiology, University of Washington, Seattle, Washington 98195, USA.
Lukehart
Sheila A
SA
D018528:ATP-Binding Cassette Transporters / Q000378:metabolism; D000890:Anti-Infective Agents / Q000378:metabolism / Q000494:pharmacology*; D002258:Carbonyl Cyanide m-Chlorophenyl Hydrazone / Q000494:pharmacology; D002352:Carrier Proteins / Q000378:metabolism; D024881:Drug Resistance, Bacterial; D006801:Humans; D048373:Treponema denticola / Q000187:drug effects* / Q000254:growth & development / Q000378:metabolism; D014475:Uncoupling Agents / Q000494:pharmacology; D023083:beta-Defensins / Q000378:metabolism / Q000494:pharmacology*
D018528:ATP-Binding Cassette Transporters; D000890:Anti-Infective Agents; D002352:Carrier Proteins; C106641:DEFB4A protein, human; D014475:Uncoupling Agents; D023083:beta-Defensins; D002258:Carbonyl Cyanide m-Chlorophenyl Hydrazone