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A comparison of pneumolysin activity and concentration in vitro and in vivo in a rabbit endophthalmitis model

Authors Sanders ME, Norcross EW, Moore III QC, Onwubiko C, King LB, Fratkin J, Marquart ME

Published 5 December 2008 Volume 2008:2(4) Pages 793—800


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Peer reviewer comments 2

Melissa E Sanders1, Erin W Norcross1, Quincy C Moore III1, Chinwendu Onwubiko1, Lauren B King1, et al

1Department of Microbiology; 2Department of Pathology, University of Mississippi Medical Center, Jackson, MS, USA

Abstract: The purpose of this study was to determine whether the in vitro activity and concentration of Streptococcus pneumoniae pneumolysin correlated to the pathogenesis of S. pneumoniae endophthalmitis. Five S. pneumoniae clinical endophthalmitis strains were grown in media to similar optical densities (OD), and extracellular milieu was tested for pneumolysin activity by hemolysis of rabbit red blood cells. Pneumolysin concentration was determined using a sandwich ELISA. Rabbit vitreous was injected with 102 colony-forming units (CFU) of 1 of 2 different strains with low hemolytic activity (n = 10 and 12 for strains 4 and 5, respectively) or 1 of 3 different strains with high hemolytic activity (n = 12 per strain). Pathogenesis of endophthalmitis infection was graded by slit lamp examination (SLE) at 24 hours post-infection. Bacteria were recovered from infected vitreous and quantitated. The SLE scores of eyes infected with strains having high hemolytic activity were significantly higher than the scores of those infected with strains having low hemolytic activity (P > 0.05). Pneumolysin concentration in vitro, however, did not correlate with hemolysis or severity of endophthalmitis. Bacterial concentrations from the vitreous infected with 4 of the strains were not significantly different (P > 0.05). These data suggest that pneumolysin hemolytic activity in vitro directly correlates to the pathogenesis of S. pneumoniae endophthalmitis. The protein concentration of pneumolysin, however, is not a reliable indicator of pneumolysin activity.

Keywords: Streptococcus pneumoniae, endophthalmitis, pneumolysin, pneumococcal

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