title: Cholesterol as treatment for pneumococcal keratitis: cholesterol-specific inhibition of pneumolysin in the cornea.
abstract: OBJECTIVE
  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 10(5) 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 log(10) 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 mug 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.
authors.affiliation: Department of Microbiology, University of Mississippi Medical Center, Jackson, Mississippi 39216, USA. mmarquart@microbio.umsmed.edu
authors.forename: Mary E
authors.initials: ME
authors.lastname: Marquart
authors.forename: Kathryn S
authors.initials: KS
authors.lastname: Monds
authors.forename: Clare C
authors.initials: CC
authors.lastname: McCormick
authors.forename: Sherrina N
authors.initials: SN
authors.lastname: Dixon
authors.forename: Melissa E
authors.initials: ME
authors.lastname: Sanders
authors.forename: Julian M
authors.initials: JM
authors.lastname: Reed
authors.forename: Larry S
authors.initials: LS
authors.lastname: McDaniel
authors.forename: Armando R
authors.initials: AR
authors.lastname: Caballero
authors.forename: Richard J
authors.initials: RJ
authors.lastname: O'Callaghan
mesh_terms: D000818:Animals; D001426:Bacterial Proteins / Q000037:antagonists & inhibitors; D002784:Cholesterol / Q000008:administration & dosage / Q000494:pharmacology / Q000627:therapeutic use*; D015169:Colony Count, Microbial; D003315:Cornea / Q000187:drug effects / Q000382:microbiology; D003320:Corneal Ulcer / Q000188:drug therapy* / Q000382:microbiology; D015818:Eye Infections, Bacterial / Q000188:drug therapy* / Q000382:microbiology; D008826:Microbial Sensitivity Tests; D009883:Ophthalmic Solutions / Q000008:administration & dosage / Q000494:pharmacology / Q000627:therapeutic use; D011008:Pneumococcal Infections / Q000188:drug therapy* / Q000382:microbiology; D011817:Rabbits; D013296:Streptococcus pneumoniae / Q000254:growth & development / Q000302:isolation & purification*; D013301:Streptolysins / Q000037:antagonists & inhibitors*
chemical_list: D001426:Bacterial Proteins; D009883:Ophthalmic Solutions; D013301:Streptolysins; C032290:plY protein, Streptococcus pneumoniae; D002784:Cholesterol