Skip to main content
PMC home page
Infection and Immunity logoLink to Infection and Immunity
. 1996 Apr;64(4):1140–1145. doi: 10.1128/iai.64.4.1140-1145.1996

Roles of autolysin and pneumolysin in middle ear inflammation caused by a type 3 Streptococcus pneumoniae strain in the chinchilla otitis media model.

K Sato 1, M K Quartey 1, C L Liebeler 1, C T Le 1, G S Giebink 1
PMCID: PMC173895  PMID: 8606070

Abstract

Streptococcus pneumoniae cell wall and pneumolysin are important contributors to pneumococcal pathogenicity in some animal models. To further explore these factors in middle ear inflammation caused by pneumococci, penicillin-induced inflammatory acceleration was studied by using three closely related pneumococcal strains: a wild-type 3 strain (WT3), its pneumolysin-negative derivative (P-1), and into autolysin-negative derivative (A-1). Both middle ears of chinchillas were inoculated with one of the three pneumococcal strains. During the first 12 h, all three strains grew in vivo at the same rate, and all three strains induced similar inflammatory cell responses in middle ear fluid (MEF). Procaine penicillin G was given as 12 h to one-half of the animals in each group, and all treated chinchillas had sterile MEF at 24 h. Penicillin significantly accelerated MEF inflammatory cell influx into WT3-and P-1-infected ears at 18 and 24 h in comparison with the rate for penicillin-treated A-1-infected ears. Inflammatory cell influx was slightly, but not significantly, greater after treatment of WT3 infection than after treatment of P-1 infection. Interleukin (IL)-1beta and IL-6, but not IL-8, concentrations in MEF at 24 h reflected the penicillin effect on MEF inflammatory cells; however, differences between treatment groups were not significant. Results suggest that pneumococcal otitis media pathogenesis is triggered principally by the inflammatory effects of intact and lytic cell wall products in the middle ear, with at most a modes additional pneumolysin effect. Investigation strategies that limit the release of these products or neutralize them warrant further investigation.

Full Text

The Full Text of this article is available as a PDF (219.5 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Alexander J. E., Lock R. A., Peeters C. C., Poolman J. T., Andrew P. W., Mitchell T. J., Hansman D., Paton J. C. Immunization of mice with pneumolysin toxoid confers a significant degree of protection against at least nine serotypes of Streptococcus pneumoniae. Infect Immun. 1994 Dec;62(12):5683–5688. doi: 10.1128/iai.62.12.5683-5688.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Berry A. M., Paton J. C., Hansman D. Effect of insertional inactivation of the genes encoding pneumolysin and autolysin on the virulence of Streptococcus pneumoniae type 3. Microb Pathog. 1992 Feb;12(2):87–93. doi: 10.1016/0882-4010(92)90111-z. [DOI] [PubMed] [Google Scholar]
  3. Berry A. M., Yother J., Briles D. E., Hansman D., Paton J. C. Reduced virulence of a defined pneumolysin-negative mutant of Streptococcus pneumoniae. Infect Immun. 1989 Jul;57(7):2037–2042. doi: 10.1128/iai.57.7.2037-2042.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bluestone C. D., Stephenson J. S., Martin L. M. Ten-year review of otitis media pathogens. Pediatr Infect Dis J. 1992 Aug;11(8 Suppl):S7–11. doi: 10.1097/00006454-199208001-00002. [DOI] [PubMed] [Google Scholar]
  5. Canafax D. M., Nonomura N., Erdmann G. R., Le C. T., Juhn S. K., Giebink G. S. Experimental animal models for studying antimicrobial pharmacokinetics in otitis media. Pharm Res. 1989 Apr;6(4):279–285. doi: 10.1023/a:1015938205892. [DOI] [PubMed] [Google Scholar]
  6. Canvin J. R., Marvin A. P., Sivakumaran M., Paton J. C., Boulnois G. J., Andrew P. W., Mitchell T. J. The role of pneumolysin and autolysin in the pathology of pneumonia and septicemia in mice infected with a type 2 pneumococcus. J Infect Dis. 1995 Jul;172(1):119–123. doi: 10.1093/infdis/172.1.119. [DOI] [PubMed] [Google Scholar]
  7. Carlsen B. D., Kawana M., Kawana C., Tomasz A., Giebink G. S. Role of the bacterial cell wall in middle ear inflammation caused by Streptococcus pneumoniae. Infect Immun. 1992 Jul;60(7):2850–2854. doi: 10.1128/iai.60.7.2850-2854.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Feldman C., Munro N. C., Jeffery P. K., Mitchell T. J., Andrew P. W., Boulnois G. J., Guerreiro D., Rohde J. A., Todd H. C., Cole P. J. Pneumolysin induces the salient histologic features of pneumococcal infection in the rat lung in vivo. Am J Respir Cell Mol Biol. 1991 Nov;5(5):416–423. doi: 10.1165/ajrcmb/5.5.416. [DOI] [PubMed] [Google Scholar]
  9. Friedland I. R., Paris M. M., Hickey S., Shelton S., Olsen K., Paton J. C., McCracken G. H. The limited role of pneumolysin in the pathogenesis of pneumococcal meningitis. J Infect Dis. 1995 Sep;172(3):805–809. doi: 10.1093/infdis/172.3.805. [DOI] [PubMed] [Google Scholar]
  10. Giebink G. S., Berzins I. K., Quie P. G. Animal models for studying pneumococcal otitis media and pneumococcal vaccine efficacy. Ann Otol Rhinol Laryngol Suppl. 1980 May-Jun;89(3 Pt 2):339–343. doi: 10.1177/00034894800890s380. [DOI] [PubMed] [Google Scholar]
  11. Giebink G. S., Juhn S. K., Weber M. L., Le C. T. The bacteriology and cytology of chronic otitis media with effusion. Pediatr Infect Dis. 1982 Mar-Apr;1(2):98–103. doi: 10.1097/00006454-198203000-00007. [DOI] [PubMed] [Google Scholar]
  12. Giebink G. S. The microbiology of otitis media. Pediatr Infect Dis J. 1989 Jan;8(1 Suppl):S18–S20. [PubMed] [Google Scholar]
  13. Himi T., Suzuki T., Kodama H., Takezawa H., Kataura A. Immunologic characteristics of cytokines in otitis media with effusion. Ann Otol Rhinol Laryngol Suppl. 1992 Oct;157:21–25. doi: 10.1177/0003489492101s1006. [DOI] [PubMed] [Google Scholar]
  14. Houldsworth S., Andrew P. W., Mitchell T. J. Pneumolysin stimulates production of tumor necrosis factor alpha and interleukin-1 beta by human mononuclear phagocytes. Infect Immun. 1994 Apr;62(4):1501–1503. doi: 10.1128/iai.62.4.1501-1503.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Johnson M. D., Fitzgerald J. E., Leonard G., Burleson J. A., Kreutzer D. L. Cytokines in experimental otitis media with effusion. Laryngoscope. 1994 Feb;104(2):191–196. doi: 10.1288/00005537-199402000-00012. [DOI] [PubMed] [Google Scholar]
  16. Johnson M. K., Boese-Marrazzo D., Pierce W. A., Jr Effects of pneumolysin on human polymorphonuclear leukocytes and platelets. Infect Immun. 1981 Oct;34(1):171–176. doi: 10.1128/iai.34.1.171-176.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Johnson M. K., Hobden J. A., O'Callaghan R. J., Hill J. M. Confirmation of the role of pneumolysin in ocular infections with Streptococcus pneumoniae. Curr Eye Res. 1992 Dec;11(12):1221–1225. doi: 10.3109/02713689208999547. [DOI] [PubMed] [Google Scholar]
  18. Juhn S. K., Garvis W. J., Lees C. J., Le C. T., Kim C. S. Determining otitis media severity from middle ear fluid analysis. Ann Otol Rhinol Laryngol Suppl. 1994 May;163:43–45. doi: 10.1177/00034894941030s512. [DOI] [PubMed] [Google Scholar]
  19. Kawana M., Kawana C., Giebink G. S. Penicillin treatment accelerates middle ear inflammation in experimental pneumococcal otitis media. Infect Immun. 1992 May;60(5):1908–1912. doi: 10.1128/iai.60.5.1908-1912.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Lee C. J., Lock R. A., Andrew P. W., Mitchell T. J., Hansman D., Paton J. C. Protection of infant mice from challenge with Streptococcus pneumoniae type 19F by immunization with a type 19F polysaccharide--pneumolysoid conjugate. Vaccine. 1994 Aug;12(10):875–878. doi: 10.1016/0264-410x(94)90028-0. [DOI] [PubMed] [Google Scholar]
  21. Leinonen M. K. Detection of Pneumococcal Capsular polysaccharide antigens by latex agglutination, counterimmunoelectrophoresis, and radioimmunoassay in middle ear exudates in acute otitis media. J Clin Microbiol. 1980 Feb;11(2):135–140. doi: 10.1128/jcm.11.2.135-140.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Lock R. A., Hansman D., Paton J. C. Comparative efficacy of autolysin and pneumolysin as immunogens protecting mice against infection by Streptococcus pneumoniae. Microb Pathog. 1992 Feb;12(2):137–143. doi: 10.1016/0882-4010(92)90116-6. [DOI] [PubMed] [Google Scholar]
  23. Maxwell K. S., Fitzgerald J. E., Burleson J. A., Leonard G., Carpenter R., Kreutzer D. L. Interleukin-8 expression in otitis media. Laryngoscope. 1994 Aug;104(8 Pt 1):989–995. doi: 10.1288/00005537-199408000-00013. [DOI] [PubMed] [Google Scholar]
  24. Moreillon P., Markiewicz Z., Nachman S., Tomasz A. Two bactericidal targets for penicillin in pneumococci: autolysis-dependent and autolysis-independent killing mechanisms. Antimicrob Agents Chemother. 1990 Jan;34(1):33–39. doi: 10.1128/aac.34.1.33. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Nonomura N., Giebink G. S., Juhn S. K., Harada T., Aeppli D. Pathophysiology of Streptococcus pneumoniae otitis media: kinetics of the middle ear biochemical and cytologic host responses. Ann Otol Rhinol Laryngol. 1991 Mar;100(3):236–243. doi: 10.1177/000348949110000313. [DOI] [PubMed] [Google Scholar]
  26. Nonomura N., Giebink G. S., Zelterman D., Harada T., Juhn S. K. Middle ear fluid lysozyme source in experimental pneumococcal otitis media. Ann Otol Rhinol Laryngol. 1991 Jul;100(7):593–596. doi: 10.1177/000348949110000715. [DOI] [PubMed] [Google Scholar]
  27. Paton J. C., Ferrante A. Inhibition of human polymorphonuclear leukocyte respiratory burst, bactericidal activity, and migration by pneumolysin. Infect Immun. 1983 Sep;41(3):1212–1216. doi: 10.1128/iai.41.3.1212-1216.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Paton J. C., Lock R. A., Hansman D. J. Effect of immunization with pneumolysin on survival time of mice challenged with Streptococcus pneumoniae. Infect Immun. 1983 May;40(2):548–552. doi: 10.1128/iai.40.2.548-552.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Rampart M., Herman A. G., Grillet B., Opdenakker G., Van Damme J. Development and application of a radioimmunoassay for interleukin-8: detection of interleukin-8 in synovial fluids from patients with inflammatory joint disease. Lab Invest. 1992 Apr;66(4):512–518. [PubMed] [Google Scholar]
  30. Rayner C. F., Jackson A. D., Rutman A., Dewar A., Mitchell T. J., Andrew P. W., Cole P. J., Wilson R. Interaction of pneumolysin-sufficient and -deficient isogenic variants of Streptococcus pneumoniae with human respiratory mucosa. Infect Immun. 1995 Feb;63(2):442–447. doi: 10.1128/iai.63.2.442-447.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Ripley-Petzoldt M. L., Giebink G. S., Juhn S. K., Aeppli D., Tomasz A., Tuomanen E. The contribution of pneumococcal cell wall to the pathogenesis of experimental otitis media. J Infect Dis. 1988 Feb;157(2):245–255. doi: 10.1093/infdis/157.2.245. [DOI] [PubMed] [Google Scholar]
  32. Rubins J. B., Charboneau D., Paton J. C., Mitchell T. J., Andrew P. W., Janoff E. N. Dual function of pneumolysin in the early pathogenesis of murine pneumococcal pneumonia. J Clin Invest. 1995 Jan;95(1):142–150. doi: 10.1172/JCI117631. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Sanchez-Puelles J. M., Ronda C., Garcia J. L., Garcia P., Lopez R., Garcia E. Searching for autolysin functions. Characterization of a pneumococcal mutant deleted in the lytA gene. Eur J Biochem. 1986 Jul 15;158(2):289–293. doi: 10.1111/j.1432-1033.1986.tb09749.x. [DOI] [PubMed] [Google Scholar]
  34. Sato K., Quartey M. K., Liebeler C. L., Giebink G. S. Timing of penicillin treatment influences the course of Streptococcus pneumoniae-induced middle ear inflammation. Antimicrob Agents Chemother. 1995 Aug;39(8):1896–1898. doi: 10.1128/aac.39.8.1896. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Takeuchi K., Maesako K., Yuta A., Sakakura Y. Interleukin-8 gene expression in middle ear effusions. Ann Otol Rhinol Laryngol. 1994 May;103(5 Pt 1):404–407. doi: 10.1177/000348949410300511. [DOI] [PubMed] [Google Scholar]
  36. Tuomanen E., Hengstler B., Rich R., Bray M. A., Zak O., Tomasz A. Nonsteroidal anti-inflammatory agents in the therapy for experimental pneumococcal meningitis. J Infect Dis. 1987 May;155(5):985–990. doi: 10.1093/infdis/155.5.985. [DOI] [PubMed] [Google Scholar]
  37. Tuomanen E., Liu H., Hengstler B., Zak O., Tomasz A. The induction of meningeal inflammation by components of the pneumococcal cell wall. J Infect Dis. 1985 May;151(5):859–868. doi: 10.1093/infdis/151.5.859. [DOI] [PubMed] [Google Scholar]
  38. Tuomanen E., Tomasz A., Hengstler B., Zak O. The relative role of bacterial cell wall and capsule in the induction of inflammation in pneumococcal meningitis. J Infect Dis. 1985 Mar;151(3):535–540. doi: 10.1093/infdis/151.3.535. [DOI] [PubMed] [Google Scholar]
  39. Van Snick J., Cayphas S., Vink A., Uyttenhove C., Coulie P. G., Rubira M. R., Simpson R. J. Purification and NH2-terminal amino acid sequence of a T-cell-derived lymphokine with growth factor activity for B-cell hybridomas. Proc Natl Acad Sci U S A. 1986 Dec;83(24):9679–9683. doi: 10.1073/pnas.83.24.9679. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Virolainen A., Salo P., Jero J., Karma P., Eskola J., Leinonen M. Comparison of PCR assay with bacterial culture for detecting Streptococcus pneumoniae in middle ear fluid of children with acute otitis media. J Clin Microbiol. 1994 Nov;32(11):2667–2670. doi: 10.1128/jcm.32.11.2667-2670.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Yellon R. F., Leonard G., Marucha P. T., Craven R., Carpenter R. J., Lehmann W. B., Burleson J. A., Kreutzer D. L. Characterization of cytokines present in middle ear effusions. Laryngoscope. 1991 Feb;101(2):165–169. doi: 10.1288/00005537-199102000-00011. [DOI] [PubMed] [Google Scholar]
  42. Yellon R. F., Leonard G., Marucha P., Sidman J., Carpenter R., Burleson J., Carlson J., Kreutzer D. Demonstration of interleukin 6 in middle ear effusions. Arch Otolaryngol Head Neck Surg. 1992 Jul;118(7):745–748. doi: 10.1001/archotol.1992.01880070075014. [DOI] [PubMed] [Google Scholar]

Articles from Infection and Immunity are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES