IOVS Infection and Immunity
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(Investigative Ophthalmology and Visual Science. 2007;48:2661-2666.)
© 2007 by The Association for Research in Vision and Ophthalmology, Inc.
DOI:  10.1167/iovs.07-0017
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Cholesterol as Treatment for Pneumococcal Keratitis: Cholesterol-Specific Inhibition of Pneumolysin in the Cornea

Mary E. Marquart, Kathryn S. Monds, Clare C. McCormick, Sherrina N. Dixon, Melissa E. Sanders, Julian M. Reed, Larry S. McDaniel, Armando R. Caballero, and Richard J. O’Callaghan

From the Department of Microbiology, University of Mississippi Medical Center, Jackson, Mississippi.

PURPOSE. The purpose of this study was to determine whether cholesterol, the host cell receptor for pneumolysin of Streptococcus pneumoniae, could effectively treat pneumococcal keratitis.

METHODS. New Zealand White rabbits were intrastromally injected with 105 colony-forming units (CFUs) of S. pneumoniae D39. Corneas were treated with topical drops of 1% cholesterol every 2 hours beginning 25 hours after infection and were examined by slit lamp microscopy 24, 36, and 48 hours after infection. Rabbits were killed, and CFUs were recovered from the corneas after the final slit lamp examination (SLE). Minimal inhibitory concentration (MIC) assays of cholesterol against bacteria were performed. Specific inhibition of pneumolysin by cholesterol in the rabbit cornea was tested by intrastromal injection of pneumolysin with or without cholesterol and was compared with cholesterol inhibition of pneumolysin in vitro using hemolysis assays with rabbit erythrocytes.

RESULTS. Corneas treated with cholesterol had significantly lower SLE scores 48 hours after infection than corneas treated with vehicle (P = 0.0015). Treated corneas also had significantly less log10 CFUs than vehicle-treated corneas (P = 0.0006). Cholesterol at a 1% concentration was bactericidal to bacteria in vitro, and lower concentrations of cholesterol were partially inhibitory in a concentration-dependent manner. Cholesterol also specifically inhibited 1 µg pneumolysin in vivo and up to 50 ng pneumolysin in vitro.

CONCLUSIONS. Topical cholesterol is an effective treatment for S. pneumoniae keratitis. Cholesterol not only inhibits pneumolysin, it is also bactericidal.




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S. N. Green, M. Sanders, Q. C. Moore III, E. W. Norcross, K. S. Monds, A. R. Caballero, L. S. McDaniel, S. A. Robinson, C. Onwubiko, R. J. O'Callaghan, et al.
Protection from Streptococcus pneumoniae Keratitis by Passive Immunization with Pneumolysin Antiserum
Invest. Ophthalmol. Vis. Sci., January 1, 2008; 49(1): 290 - 294.
[Abstract] [Full Text] [PDF]


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