Pathogenesis of A Clinical Ocular Strain of Streptococcus pneumoniae and the Interaction of Pneumolysin with Corneal Cells.

Streptococcus pneumoniae is an important cause of bacterial keratitis, an infectious disease of the cornea. This study aimed to determine the importance of pneumolysin (PLY), a pneumococcal virulence factor, in keratitis using a clinical keratitis isolate (K1263) and its isogenic mutant deficient in PLY (K1263ΔPLY) and determine the effect of these strains on primary rabbit corneal epithelial (RCE) cells. Each strain was injected into the corneal stromas of rabbits, clinical examinations were performed, and the recovered bacterial loads were determined. Bacterial extracts were exposed to RCE cells, and morphology and viability were assessed. The mutant strain deficient in PLY, K1263ΔPLY, caused significantly lower ocular disease scores than the parent strain (K1263), although a higher bacterial load was recovered from corneas infected with the mutant strain. Histological examination showed increased inflammatory cells in the anterior chamber and increased edema in eyes infected with the parent strain. RCE cells exposed to the parent strain had significantly decreased cell viability and showed increased evidence of cellular damage. This study confirms that in a strain that can cause clinical keratitis, PLY is a significant cause of the damage associated with pneumococcal keratitis. It also shows for the first time that the results from an in vitro model using RCE cells correlates with in vivo results thereby establishing a less invasive way to study the mechanisms of pneumococcal keratitis.