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Relative roles of pneumolysin and hydrogen peroxide from Streptococcus pneumoniae in inhibition of ependymal ciliary beat frequency

By Robert A. Hirst, Kulvinder S. Sikand, Andrew Rutman, Timothy J. Mitchell, Peter W. Andrew and Christopher L. O'Callaghan


Copyright © 2000, American Society for Microbiology. All Rights Reserved. Also available from the publisher website at http://iai.asm.org/Ciliated ependymal cells line the ventricular system of the brain and the cerebral aqueducts. This study characterizes the relative roles of pneumolysin and hydrogen peroxide (H2O2) in pneumococcal meningitis, using the in vitro ependymal ciliary beat frequency (CBF) as an indicator of toxicity. We have developed an ex vivo model to examine the ependymal surface of the brain slices cut from the fourth ventricle. The ependymal cells had cilia beating at a frequency of between 38 and 44Hz. D39 (wild-type) and PLN-A (pneumolysinnegative) pneumococci at 108 CFU/ml both caused ciliary slowing. Catalase protected against PLN-A-induced ciliary slowing but afforded little protection from D39. Lysed PLN-A did not reduce CBF, whereas lysed D39 caused rapid ciliary stasis. There was no effect of catalase, penicillin, or catalase plus penicillin on the CBF. H2O2 at a concentration as low as 100 mM caused ciliary stasis, and this effect was abolished by coincubation with catalase. An additive inhibition of CBF was demonstrated using a combination of both toxins. A significant inhibition of CBF at between 30 and 120 min was demonstrated with both toxins compared with either H2O2 (10 mM) or pneumolysin (1 HU/ml) alone. D39 released equivalent levels of H2O2 to those released by PLN-A, and these concentrations were sufficient to cause ciliary stasis. The brain slices did not produce H2O2, and in\ud the presence of 108 CFU of D39 or PLN-A per ml there was no detectable bacterially induced increase of H2O2 release from the brain slice. Coincubation with catalase converted the H2O2 produced by the pneumococci to H2O. Penicillin-induced lysis of bacteria dramatically reduced H2O2 production. The hemolytic activity released\ud from D39 was sufficient to cause rapid ciliary stasis, and there was no detectable release of hemolytic activity from the pneumolysin-negative PLN-A. These data demonstrate that D39 bacteria released pneumolysin, which caused rapid ciliary stasis. D39 also released H2O2, which contributed to the toxicity, but this was\ud masked by the more severe effects of pneumolysin. H2O2 released from intact PLN-A was sufficient to cause rapid ciliary stasis, and catalase protected against H2O2-induced cell toxicity, indicating a role for H2O2 in the response. There is also a slight additive effect of pneumolysin and H2O2 on ependymal toxicity; however, the precise mechanism of action and the role of these toxins in pathogenesis remain unclear

Publisher: American Society for Microbiology
Year: 2000
OAI identifier: oai:lra.le.ac.uk:2381/2684

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  2. (1990). Bacterial counts in cerebrospinal fluid of children with meningitis. doi
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  4. (1991). Cellular injury by oxidants. doi
  5. (1988). Ciliary defects: cell biology and clinical perspectives.
  6. (1985). Cyclic AMP and calcium in the differential control of mytillus gill cilia. doi
  7. Effect of pneumolysin on rat brain ciliary function: comparison of brain slices with cultured ependymal cells. doi
  8. (1985). Effects of ampicillin and corticosteroids on brain water content, CSF pressure and CSF lactate in experimental pneumococcal meningitis. doi
  9. (1991). Effects of P. aeruginosa-derived bacterial products on tracheal ciliary function: the role of O2 radicals.
  10. (1989). Expression of the pneumolysin gene in Escherichia coli: rapid purification and biological properties. doi
  11. (1981). Hydrogen peroxide and superoxide release by alveolar macrophages from normal and BCG-vaccinated guinea pigs after intravenous challenge with mycobacterium tuberculosis.
  12. (1999). Identification of a neural stem cell in the adult mammalian central nervous system. doi
  13. (1993). Identification of hydrogen peroxide as a Streptococcus pneumoniae toxin for rat alveolar epithelial cells.
  14. (1995). Interaction of pneumolysin-sufficient and deficient isogenic variants of Streptococcus pneumoniae with human respiratory mucosa.
  15. (1959). Lactic oxidase of the pneumococcus.
  16. (1992). Mechanism of hydrogen peroxide-induced inhibition of sheep airway cilia. doi
  17. (1991). Mechanisms of endothelial cell killing by H2O2 or products of activated neutrophils. doi
  18. (1993). Molecular analysis of the pathogenicity of Streptococcus pneumoniae: the role of pneumococcal proteins. doi
  19. (1994). Molecular cloning and sequence analysis of the gene encoding the H2O2-forming NADH oxidase from Streptococcus mutans. doi
  20. (1986). Oxidant-mediated ciliary dysfunction. Possible role in airway disease. doi
  21. (1995). Pathogenesis of pneumococcal infection. doi
  22. (1984). Pneumococcal meningitis: late neurological sequelae and features of prognostic impact. doi
  23. (1994). Polymorphonuclear leukocyte-generated oxygen metabolites decrease beat frequency of human respiratory cilia. Lung 172:215–222. doi
  24. (1994). Protective effects of a cyclic nitrone antioxidant in animal models of endotoxic shock and chronic bacteraemia.
  25. (1996). Pyruvate oxidase, as a determinant of virulence in Streptococcus pneumoniae. doi
  26. (1996). Reactive oxygen intermediates contribute to necrotic and apoptotic neuronal injury in an infant rat model of bacterial meningitis due to group B streptococci. doi
  27. (1989). Reduced virulence of a defined pneumolysin-negative mutant of Streptococcus pneumoniae.
  28. (1999). The effect of the pneumococcal toxin, pneumolysin on brain ependymal cilia. doi
  29. (1995). The ependyma: a protective barrier between brain and cerebrospinal fluid. doi

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