Abstract
Purpose: :
To examine the cytopathic and immunological changes associated with live infection of primary human corneal fibroblasts with Pseudomonas aeruginosa.
Methods: :
Human corneal fibroblasts (CF) were extracted from clinical samples, cultured to confluence in vitro and infected with various doses of Pseudomonas aeruginosa PAO-1. Bacterial association and invasion were quantified at 3, 6 and 9 hours and host cell death and cytokine profiles were assessed at 24 hours post challenge.
Results: :
PAO-1 adhered to human CF in a dose and time-dependant manner and cytotoxic effects of bacterial interactions were observed within 9h. Preliminary data shows that constitutive levels of IL-8 are reduced with increasing infective dose, which reflects the pattern of cytotoxicity.
Conclusions: :
We established an in vitro model of microbial keratitis and demonstrated the direct adherence of Pseudomonas aeruginosa to CF and the concurrent induction of host cell cytoxicity. The innate immune recognition of bacteria results in a decrease in resting levels of IL-8 in a dose dependent manner.
Keywords: cornea: basic science • microbial pathogenesis: experimental studies • inflammation