title: Type III Secretion-Dependent Sensitivity of Escherichia coli O157 to Specific Ketolides.
abstract: A subset of Gram-negative bacterial pathogens uses a type III secretion system (T3SS) to open up a conduit into eukaryotic cells in order to inject effector proteins. These modulate pathways to enhance bacterial colonization. In this study, we screened established bioactive compounds for any that could repress T3SS expression in enterohemorrhagic Escherichia coli (EHEC) O157. The ketolides telithromycin and, subsequently, solithromycin both demonstrated repressive effects on expression of the bacterial T3SS at sub-MICs, leading to significant reductions in bacterial binding and actin-rich pedestal formation on epithelial cells. Preincubation of epithelial cells with solithromycin resulted in significantly less attachment of E. coli O157. Moreover, bacteria expressing the T3SS were more susceptible to solithromycin, and there was significant preferential killing of E. coli O157 bacteria when they were added to epithelial cells that had been preexposed to the ketolide. This killing was dependent on expression of the T3SS. Taken together, this research indicates that the ketolide that has accumulated in epithelial cells may traffic back into the bacteria via the T3SS. Considering that neither ketolide induces the SOS response, nontoxic members of this class of antibiotics, such as solithromycin, should be considered for future testing and trials evaluating their use for treatment of EHEC infections. These antibiotics may also have broader significance for treating infections caused by other pathogenic bacteria, including intracellular bacteria, that express a T3SS.
authors.affiliation: Division of Immunity and Infection, The Roslin Institute and R(D)SVS, The University of Edinburgh, Easter Bush, Midlothian, United Kingdom Laboratory of Pathogenesis and Immunology of Infectious Diseases, IMEX-CONICET, National Academy of Medicine, Buenos Aires, Argentina.
authors.forename: Romina J
authors.initials: RJ
authors.lastname: Fernandez-Brando
authors.affiliation: Division of Immunity and Infection, The Roslin Institute and R(D)SVS, The University of Edinburgh, Easter Bush, Midlothian, United Kingdom.
authors.forename: Nao
authors.initials: N
authors.lastname: Yamaguchi
authors.affiliation: Division of Immunity and Infection, The Roslin Institute and R(D)SVS, The University of Edinburgh, Easter Bush, Midlothian, United Kingdom Faculty of Veterinary Medicine, Kafrelsheikh University, Kafrel-Sheikh, Egypt.
authors.forename: Amin
authors.initials: A
authors.lastname: Tahoun
authors.affiliation: Division of Immunity and Infection, The Roslin Institute and R(D)SVS, The University of Edinburgh, Easter Bush, Midlothian, United Kingdom.
authors.forename: Sean P
authors.initials: SP
authors.lastname: McAteer
authors.affiliation: IMPACT Facility, Centre for Integrative Physiology, The University of Edinburgh, Edinburgh, United Kingdom.
authors.forename: Trudi
authors.initials: T
authors.lastname: Gillespie
authors.affiliation: Division of Immunity and Infection, The Roslin Institute and R(D)SVS, The University of Edinburgh, Easter Bush, Midlothian, United Kingdom State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, Xiamen University, Xiamen, Fujian, People's Republic of China.
authors.forename: Dai
authors.initials: D
authors.lastname: Wang
authors.affiliation: Division of Immunity and Infection, The Roslin Institute and R(D)SVS, The University of Edinburgh, Easter Bush, Midlothian, United Kingdom.
authors.forename: Sally A
authors.initials: SA
authors.lastname: Argyle
authors.affiliation: Laboratory of Pathogenesis and Immunology of Infectious Diseases, IMEX-CONICET, National Academy of Medicine, Buenos Aires, Argentina.
authors.forename: Marina S
authors.initials: MS
authors.lastname: Palermo
authors.affiliation: Division of Immunity and Infection, The Roslin Institute and R(D)SVS, The University of Edinburgh, Easter Bush, Midlothian, United Kingdom dgally@ed.ac.uk.
authors.forename: David L
authors.initials: DL
authors.lastname: Gally
mesh_terms: D000818:Animals; D000900:Anti-Bacterial Agents / Q000737:chemistry / Q000494:pharmacology*; D001422:Bacterial Adhesion / Q000187:drug effects; D018938:Caco-2 Cells; D002417:Cattle; D002460:Cell Line; D055808:Drug Discovery; D004847:Epithelial Cells / Q000187:drug effects / Q000382:microbiology; D019453:Escherichia coli O157 / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D057166:High-Throughput Screening Assays; D006801:Humans; D048628:Ketolides / Q000737:chemistry / Q000494:pharmacology*; D018942:Macrolides / Q000737:chemistry / Q000494:pharmacology*; D008826:Microbial Sensitivity Tests; D020545:Respiratory Mucosa / Q000187:drug effects / Q000382:microbiology; D054852:Small Molecule Libraries / Q000494:pharmacology*; D014230:Triazoles / Q000737:chemistry / Q000494:pharmacology*; D000069296:Type III Secretion Systems / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism
chemical_list: D000900:Anti-Bacterial Agents; D048628:Ketolides; D018942:Macrolides; D054852:Small Molecule Libraries; D014230:Triazoles; D000069296:Type III Secretion Systems; C547755:solithromycin; C106791:telithromycin