PURPOSE: To determine the pathogenic role of gamma- and alpha-toxin in a rabbit model of Staphylococcus aureus keratitis. METHODS: S. aureus strains Newman (expressing gamma-toxin), Newman Delta(hlg) (deficient in gamma-toxin), Newman Delta(hlg)/pCU1 hlg(+) (chromosomal gamma-toxin-deficient mutant rescued by a plasmid encoding gamma-toxin), and Newman Delta(hla) (alpha-toxin-deficient) were intrastromally injected into rabbit corneas. Eyes were scored by slit lamp examination (SLE), and bacterial colony-forming units (CFU) per cornea were determined at 15, 20, and 25 hours after infection. Histologic examination of corneas was performed. Rabbits were immunized against alpha-toxin and subsequently challenged with S. aureus strain Newman. Western blot analyses of culture supernatants were performed to detect alpha-toxin production. RESULTS: All strains grew equivalently, producing approximately 7 log CFU per cornea at 25 hours after infection. SLE scores at 20 and 25 hours after infection revealed that strains Newman Delta(hlg) and Newman Delta(hla), although virulent, caused significantly less ocular damage and inflammation than their parent or the gamma-toxin genetically rescued strain (P <or= 0.0006). Histologic and SLEs revealed that all strains except Newman Delta(hla) produced corneal erosions. Rabbits immunized actively or passively to alpha-toxin had reduced SLE scores (P <or= 0.0003 and P <or= 0.0033, respectively) and no epithelial erosions when infected with strain Newman. Western blot analysis demonstrated that strains Newman and Newman Delta(hlg), but not Newman Delta(hla), produced alpha-toxin. CONCLUSIONS: These results illustrate that the virulence of strain Newman involves both alpha- and gamma-toxin, with alpha-toxin mediating corneal epithelial erosions. An additional uncharacterized toxin could also be active in damaging the cornea.
PURPOSE: To determine the pathogenic role of gamma- and alpha-toxin in a rabbit model of Staphylococcus aureus keratitis. METHODS:S. aureus strains Newman (expressing gamma-toxin), Newman Delta(hlg) (deficient in gamma-toxin), Newman Delta(hlg)/pCU1 hlg(+) (chromosomal gamma-toxin-deficient mutant rescued by a plasmid encoding gamma-toxin), and Newman Delta(hla) (alpha-toxin-deficient) were intrastromally injected into rabbit corneas. Eyes were scored by slit lamp examination (SLE), and bacterial colony-forming units (CFU) per cornea were determined at 15, 20, and 25 hours after infection. Histologic examination of corneas was performed. Rabbits were immunized against alpha-toxin and subsequently challenged with S. aureus strain Newman. Western blot analyses of culture supernatants were performed to detect alpha-toxin production. RESULTS: All strains grew equivalently, producing approximately 7 log CFU per cornea at 25 hours after infection. SLE scores at 20 and 25 hours after infection revealed that strains Newman Delta(hlg) and Newman Delta(hla), although virulent, caused significantly less ocular damage and inflammation than their parent or the gamma-toxin genetically rescued strain (P <or= 0.0006). Histologic and SLEs revealed that all strains except Newman Delta(hla) produced corneal erosions. Rabbits immunized actively or passively to alpha-toxin had reduced SLE scores (P <or= 0.0003 and P <or= 0.0033, respectively) and no epithelial erosions when infected with strain Newman. Western blot analysis demonstrated that strains Newman and Newman Delta(hlg), but not Newman Delta(hla), produced alpha-toxin. CONCLUSIONS: These results illustrate that the virulence of strain Newman involves both alpha- and gamma-toxin, with alpha-toxin mediating corneal epithelial erosions. An additional uncharacterized toxin could also be active in damaging the cornea.
Authors: B Saïd-Salim; P M Dunman; F M McAleese; D Macapagal; E Murphy; P J McNamara; S Arvidson; T J Foster; S J Projan; B N Kreiswirth Journal: J Bacteriol Date: 2003-01 Impact factor: 3.490
Authors: Aihua Tang; Mary E Marquart; Jonathan D Fratkin; Clare C McCormick; Armando R Caballero; Hattie P Gatlin; Richard J O'Callaghan Journal: Invest Ophthalmol Vis Sci Date: 2009-02-28 Impact factor: 4.799