In vitro screens for quorum sensing inhibitors and in vivo confirmation of their effect.
This article will introduce the reader to protocols intended for (i) identification of quorum sensing (QS) inhibitors (QSIs), (ii) characterization of these compounds in vitro and (iii) evaluation of these compounds in animal models. Traditional antimicrobial drugs are designed against planktonic bacteria and not against bacterial biofilms. In biofilms, bacteria are highly resistant to otherwise lethal treatments and they communicate with each other, thus enabling coordinated group behavior. For many years, we have focused on interference with cell to cell communication, also known as QS, with the aim of disabling the expression of virulence and reduction of antibiotic tolerance. Here we present protocols for screening and testing for acyl-homoserine lactone (AHL)-dependent QS inhibition. We also present protocols for the in vivo validation of QSIs as possible drug candidates. The presented methods allow the evaluation of QS inhibition by a potential drug candidate within 2-3 weeks.
Department of International Health, Immunology and Microbiology, Faculty of Health Sciences, University of Copenhagen, Copenhagen, Denmark.
Thomas
T
Bjarnsholt
Maria
M
van Gennip
Tim H
TH
Jakobsen
Louise D
LD
Christensen
Peter Ø
PØ
Jensen
Michael
M
Givskov
D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D001419:Bacteria / Q000187:drug effects; D001431:Bacteriological Techniques; D004195:Disease Models, Animal; D024881:Drug Resistance, Bacterial / Q000235:genetics; D004927:Escherichia coli Infections / Q000188:drug therapy; D006090:Gram-Negative Bacteria / Q000187:drug effects / Q000235:genetics; D006801:Humans; D008171:Lung Diseases / Q000382:microbiology; D018613:Microscopy, Confocal / Q000379:methods; D008855:Microscopy, Electron, Scanning / Q000379:methods; D023421:Models, Animal; D053038:Quorum Sensing / Q000187:drug effects* / Q000235:genetics
D000900:Anti-Bacterial Agents