title: Influence of the protein kinase C activator phorbol myristate acetate on the intracellular activity of antibiotics against hemin- and menadione-auxotrophic small-colony variant mutants of Staphylococcus aureus and their wild-type parental strain in human THP-1 cells.
abstract: In a previous study (L. G. Garcia et al., Antimicrob. Agents Chemother. 56:3700-3711, 2012), we evaluated the intracellular fate of menD and hemB mutants (corresponding to menadione- and hemin-dependent small-colony variants, respectively) of the parental COL methicillin-resistant Staphylococcus aureus strain and the pharmacodynamic profile of the intracellular activity of a series of antibiotics in human THP-1 monocytes. We have now examined the phagocytosis and intracellular persistence of the same strains in THP-1 cells activated by phorbol 12-myristate 13-acetate (PMA) and measured the intracellular activity of gentamicin, moxifloxacin, and oritavancin in these cells. Postphagocytosis intracellular counts and intracellular survival were lower in PMA-activated cells, probably due to their higher killing capacities. Gentamicin and moxifloxacin showed a 5- to 7-fold higher potency (lower static concentrations) against the parental strain, its hemB mutant, and the genetically complemented strain in PMA-activated cells and against the menD strain in both activated and nonactivated cells. This effect was inhibited when cells were incubated with N-acetylcysteine (a scavenger of oxidant species). In parallel, we observed that the MICs of these drugs were markedly reduced if bacteria had been preexposed to H(2)O(2). In contrast, the intracellular potency of oritavancin was not different in activated and nonactivated cells and was not decreased by the addition of N-acetylcysteine, regardless of the phenotype of the strains. The oritavancin MIC was also unaffected by preincubation of the bacteria with H(2)O(2). Thus, activation of THP-1 cells by PMA may increase the intracellular potency of certain antibiotics (probably due to synergy with reactive oxygen species), but this effect cannot be generalized to all antibiotics.
authors.affiliation: Pharmacologie cellulaire et moléculaire and Louvain Drug Research Institute, Université catholique de Louvain, Brussels, Belgium.
authors.forename: Laetitia G
authors.initials: LG
authors.lastname: Garcia
authors.forename: Sandrine
authors.initials: S
authors.lastname: Lemaire
authors.forename: Barbara C
authors.initials: BC
authors.lastname: Kahl
authors.forename: Karsten
authors.initials: K
authors.lastname: Becker
authors.forename: Richard A
authors.initials: RA
authors.lastname: Proctor
authors.forename: Paul M
authors.initials: PM
authors.lastname: Tulkens
authors.forename: Françoise
authors.initials: F
authors.lastname: Van Bambeke
mesh_terms: D000111:Acetylcysteine / Q000494:pharmacology; D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D002454:Cell Differentiation / Q000187:drug effects; D002460:Cell Line; D024881:Drug Resistance, Bacterial; D020536:Enzyme Activators / Q000494:pharmacology*; D005839:Gentamicins / Q000627:therapeutic use; D006427:Hemin / Q000235:genetics*; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology; D008826:Microbial Sensitivity Tests; D009000:Monocytes / Q000382:microbiology; D009154:Mutation / Q000235:genetics / Q000502:physiology*; D010587:Phagocytosis / Q000187:drug effects; D011493:Protein Kinase C / Q000378:metabolism*; D013211:Staphylococcus aureus / Q000187:drug effects* / Q000235:genetics; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology*; D024483:Vitamin K 3 / Q000378:metabolism*
chemical_list: D000900:Anti-Bacterial Agents; D020536:Enzyme Activators; D005839:Gentamicins; D024483:Vitamin K 3; D006427:Hemin; D006861:Hydrogen Peroxide; D011493:Protein Kinase C; D013755:Tetradecanoylphorbol Acetate; D000111:Acetylcysteine