Effects of subinhibitory concentrations of antibiotics on colonization factor expression by moxifloxacin-susceptible and moxifloxacin-resistant Clostridium difficile strains.
Recent outbreaks of Clostridium difficile infection have been related to the emergence of the NAP1/027 epidemic strain. This strain demonstrates increased virulence and resistance to the C-8-methoxyfluoroquinolones gatifloxacin and moxifloxacin. These antibiotics have been implicated as major C. difficile infection-inducing agents. We investigated by real-time reverse transcription-PCR the impact of subinhibitory concentrations of ampicillin, clindamycin, ofloxacin, and moxifloxacin on the expression of genes encoding three colonization factors, the protease Cwp84, the high-molecular-weight S-layer protein, and the fibronectin-binding protein Fbp68. We have previously shown in six non-NAP1/027 moxifloxacin-susceptible strains that the presence of ampicillin or clindamycin induced an upregulation of these genes, whereas the presence of fluoroquinolones did not. The objective of this study was to analyze the expression of these genes under the same conditions in four NAP1/027 strains, one moxifloxacin susceptible and three moxifloxacin resistant. Two in vitro-selected moxifloxacin-resistant mutants were also analyzed. Moxifloxacin resistance was associated with the Thr82-->Ile substitution in GyrA in all but one of the moxifloxacin-resistant strains. The expression of cwp84 and slpA was strongly increased after culture with ampicillin or clindamycin in NAP1/027 strains. Interestingly, after culture with fluoroquinolones, the expression of cwp84 and slpA was only increased in four moxifloxacin-resistant strains, including the NAP1/027 strains and one of the in vitro-selected mutants. The overexpression of cwp84 was correlated with increased production of the protease Cwp84. The historical NAP1/027 moxifloxacin-susceptible strain and its mutant appear to be differently regulated by fluoroquinolones. Overall, fluoroquinolones appear to favor the expression of some colonization factor-encoding genes in resistant C. difficile strains. The fluoroquinolone resistance of the NAP1/027 epidemic strains could be considered an ecological advantage. This could also increase their colonization fitness and promote the infection.
Denève
Cécile
C
EA 4043, Université Paris-Sud 11, Faculté de Pharmacie, Châtenay-Malabry Cedex, France.
Bouttier
Sylvie
S
Dupuy
Bruno
B
Barbut
Frédéric
F
Collignon
Anne
A
Janoir
Claire
C
D000667:Ampicillin / Q000494:pharmacology; D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D001372:Aza Compounds / Q000494:pharmacology; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D002981:Clindamycin / Q000494:pharmacology; D016360:Clostridioides difficile / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism*; D003546:Cysteine Endopeptidases / Q000235:genetics; D027081:DNA Gyrase / Q000235:genetics; D024841:Fluoroquinolones; D016368:Frameshift Mutation; D015151:Immunoblotting; D008826:Microbial Sensitivity Tests; D000077266:Moxifloxacin; D015242:Ofloxacin / Q000494:pharmacology; D016133:Polymerase Chain Reaction; D011804:Quinolines / Q000494:pharmacology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017384:Sequence Deletion
D000900:Anti-Bacterial Agents; D001372:Aza Compounds; D001426:Bacterial Proteins; D024841:Fluoroquinolones; D011804:Quinolines; C070683:surface layer protein A, Bacteria; D002981:Clindamycin; D000667:Ampicillin; D015242:Ofloxacin; C524258:Cwp84 protein, Clostridium difficile; D003546:Cysteine Endopeptidases; D027081:DNA Gyrase; D000077266:Moxifloxacin