title: Enhanced production of phospholipase C and perfringolysin O (alpha and theta toxins) in a gatifloxacin-resistant strain of Clostridium perfringens.
abstract: Clostridium perfringens-induced gas gangrene is mediated by potent extracellular toxins, especially alpha toxin (a phospholipase C [PLC]) and theta toxin (perfringolysin O [PFO], a thiol-activated cytolysin); and antibiotic-induced suppression of toxin synthesis is an important clinical goal. The production of PLC and PFO by a gatifloxacin-induced, fluoroquinolone-resistant mutant strain of C. perfringens, strain 10G, carrying a stable mutation in DNA gyrase was compared with that of the wild-type (WT) parent strain. Zymography (with sheep red blood cell and egg yolk overlays) and time course analysis [with hydrolysis of egg yolk lecithin and O-(4 nitrophenyl-phosphoryl)choline] demonstrated that strain 10G produced more PLC and PFO than the WT strain. Increased toxin production in strain 10G was not related either to differences in growth characteristics between the wild-type and the mutant strain or to nonsynonymous polymorphisms in PLC, PFO, or their known regulatory proteins. Increased PLC and PFO production by strain 10G was associated with increased cytotoxic activity for HT-29 human adenocarcinoma cells and with increased platelet-neutrophil aggregate formation. Four other gatifloxacin-induced gyrase mutants did not show increased toxin production, suggesting that gatifloxacin resistance was not always associated with increased toxin production in all strains of C. perfringens. This is the first report of increased toxin production in a fluoroquinolone-resistant strain of C. perfringens.
authors.affiliation: Division of Microbiology, National Center for Toxicological Research, 3900 NCTR Dr., Jefferson, AR 72079, USA. Fatemeh.Rafii@fda.hhs.gov
authors.forename: Fatemeh
authors.initials: F
authors.lastname: Rafii
authors.forename: Miseon
authors.initials: M
authors.lastname: Park
authors.forename: Amy E
authors.initials: AE
authors.lastname: Bryant
authors.forename: Shemedia J
authors.initials: SJ
authors.lastname: Johnson
authors.forename: Robert D
authors.initials: RD
authors.lastname: Wagner
mesh_terms: D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D001427:Bacterial Toxins / Q000096:biosynthesis* / Q000235:genetics; D045744:Cell Line, Tumor; D003016:Clostridium perfringens / Q000187:drug effects* / Q000201:enzymology / Q000254:growth & development / Q000378:metabolism; D024881:Drug Resistance, Bacterial / Q000235:genetics; D024841:Fluoroquinolones / Q000494:pharmacology*; D000077734:Gatifloxacin; D006460:Hemolysin Proteins / Q000096:biosynthesis* / Q000235:genetics; D006801:Humans; D009154:Mutation; D010738:Type C Phospholipases / Q000096:biosynthesis* / Q000235:genetics; D015854:Up-Regulation
chemical_list: D000900:Anti-Bacterial Agents; D001427:Bacterial Toxins; D024841:Fluoroquinolones; D006460:Hemolysin Proteins; C020452:Clostridium perfringens theta-toxin; D010738:Type C Phospholipases; D000077734:Gatifloxacin