The Effect of Transformation on the Virulence of Streptococcus pneumoniae

  • Zhang Xue-Mei (Faculty of Laboratory Medicine, Chongqing University of Medical Sciences) ;
  • Yin Yi-Bing (Faculty of Laboratory Medicine, Chongqing University of Medical Sciences) ;
  • Zhu Dan (Faculty of Laboratory Medicine, Chongqing University of Medical Sciences) ;
  • Chen Bao-De (Faculty of Laboratory Medicine, Chongqing University of Medical Sciences) ;
  • Luo Jin-Yong (Faculty of Laboratory Medicine, Chongqing University of Medical Sciences) ;
  • Deng Vi-Ping (Faculty of Laboratory Medicine, Chongqing University of Medical Sciences) ;
  • Liu Ming-Fang (Faculty of Laboratory Medicine, Chongqing University of Medical Sciences) ;
  • Chen Shu-Hui (Faculty of Laboratory Medicine, Chongqing University of Medical Sciences) ;
  • Meng Jiang-Ping (Faculty of Laboratory Medicine, Chongqing University of Medical Sciences) ;
  • Lan Kai (Faculty of Laboratory Medicine, Chongqing University of Medical Sciences) ;
  • Huang Yuan-Shuai (Faculty of Laboratory Medicine, Chongqing University of Medical Sciences) ;
  • Kang Ge-Fei (Faculty of Laboratory Medicine, Chongqing University of Medical Sciences)
  • Published : 2005.08.01

Abstract

Although pneumococcus is one of the most frequently encountered opportunistic pathogen in the world, the mechanisms responsible for its infectiveness have not yet been fully understood. In this paper, we have attempted to characterize the effects of pneumococcal transformation on the pathogenesis of the organism. We constructed three transformation-deficient pneumococcal strains, which were designated as Nos. 1d, 2d, and 22d. The construction of these altered strains was achieved via the insertion of the inactivated gene, comE, to strains 1, 2 and 22. We then conducted a comparison between the virulence of the transformation-deficient strains and that of the wild-type strains, via an evaluation of the ability of each strain to adhere to endothelial cells, and also assessed psaA mRNA expression, and the survival of hosts after bacterial challenge. Compared to what was observed with the wild-type strains, our results indicated that the ability of all of the transformation-deficient strains to adhere to the ECV304 cells had been significantly reduced (p < 0.05), the expression of psaA mRNA was reduced significantly (p < 0.05) in strains 2d and 22d, and the median survival time of mice infected with strains Id and 2d was increased significantly after intraperitoneal bacterial challenge (p < 0.05). The results of our study also clearly indicated that transformation exerts significant effects on the virulence characteristics of S. pneumoniae, although the degree to which this effect is noted appears to depend primarily on the genetic background of the bacteria.

Keywords

References

  1. Austrian, R. 1981. Some observations on the pneumococcus and on the current status of pneumococcal disease and its prevention. Rev. Infect. Dis. 3, 1-17 https://doi.org/10.1093/clinids/3.1.1
  2. Auzat, I., S. Chapuy-Regaud, D. Dos Santos, I. Le Thomas, G. Le Bras, D. Onnuyighy, J.-R. Garel, J. Paton, and M.C. Trombe. 1999. The NADH oxidase of Streptococcus pneumoniae, its role in competence and virulence. Mol. Microbiol. 34, 1018- 1028 https://doi.org/10.1046/j.1365-2958.1999.01663.x
  3. Balachandran, P., A. Brooks-Walter, A. Virolainen-Julkunen, S.K. Hollingshead, and D.E. Briles. 2002. Role of pneumococcal surface protein C in nasopharyngeal carriage and pneumonia and its ability to elicit protection against carriage of Streptococcus pneumoniae. Infect. Immun. 70, 2526-2534 https://doi.org/10.1128/IAI.70.5.2526-2534.2002
  4. Bartilson, M., A. Marra, J. Christine, J.S. Asundi, W.P. Schneider, and A.E. Hromockyj. 2001. Differential fluorescence induction reveals Streptococcus pneumoniae loci regulated by competence stimulatory peptide. Mol. Microbiol. 39, 126-135 https://doi.org/10.1046/j.1365-2958.2001.02218.x
  5. Berry, A.M., R.A. Lock, J.C. Paton. 1996. Cloning and characterization of nanB, a sencond Streptococcus pneumoniae neuraminidase gene, and purification of the NanB enzyme from recombinant Escherichia coli. J. Bacteriol. 178, 4854-4860 https://doi.org/10.1128/jb.178.16.4854-4860.1996
  6. Berry, A.M. and J.C. Paton. 1996. Sequence Heterogeneity of PsaA, a 37-Kilodalton Putative Adhesin Essential for Virulence of Streptococcus pneumoniae. Infect. Immun. 64, 5255-5262
  7. Briles, D.E., S. Hollingshead, A. Brooks-Walter, G.S. Nabors, L. Ferguson, M. Schilling, S. Gravenstein, P. Braun, J. and King, A. Swift. 2000. The potential to use PspA and other pneumococcal proteins to elicit protection against pneumococcal infection. Vaccine 18, 1707-1711 https://doi.org/10.1016/S0264-410X(99)00511-3
  8. Chapuy-Regaud, S., A.D. Ogunniyi, N. Diallo, Y. Huet, J.F. Desnottes, J.C. Paton, S. Escaich, and M.C. Trombe. 2003. RegR, a Global LacI/GalR Family Regulator, Modulates Virulence and Competence in Streptococcus pneumoniae. Infect. Immun. 71, 2615-2625 https://doi.org/10.1128/IAI.71.5.2615-2625.2003
  9. Cheng, Q., E.A. Campbell, A.M. Naughton, S. Johnson, and H.R. Masure. 1997. The com locus controls genetic transformation in Streptococcus pneumoniae. Mol. Microbiol. 23, 683-692 https://doi.org/10.1046/j.1365-2958.1997.2481617.x
  10. Cundell, D.R., N.P. Gerard, C. Gerard, I. Idanpaan-Heikkila, and E.I. Tuomanen. 1995. Streptococcus pneumoniae anchor to activate human cells by the receptor for platelet-activating factor. Nature 377, 435-438 https://doi.org/10.1038/377435a0
  11. Dowson, C.G., V. Barcus, S. King, P. Pickerill, A. Whatmore, and M. Yeo. 1997. Horizontal gene transfer and the evolution of resistance and virulence determinants in Streptococcus. Soc. Appl. Bacteriol. Symp. Ser. 26, 42S-51S
  12. Echenique, J.R. and M.C. Trombe. 2001. Trombe. Competence modulation by the NADH oxidase of Streptococcus pneumoniae involves signal transduction. J. Bacteriol. 183, 768-772 https://doi.org/10.1128/JB.183.2.768-772.2001
  13. Geelen, S., C. Bhattacharyya, and E. Tuomanen. 1993. The cell wall mediates pneumococcal attachment to and cytopathology in human endothelial cells. Infect. Immun. 61, 1538-1543
  14. Hammerschmidt, S., S.R. Talay, P. Brandtzaeg, and G.S. Chhatwal. 1997. SpsA, a novel pnumococcal surface protein with specific binding to secretory immunoglobulin A and secretory component. Mol. Microbiol. 25, 1113-1124 https://doi.org/10.1046/j.1365-2958.1997.5391899.x
  15. Havarstein, L.S., G. Coomaraswamy, and D.A. Morrison. 1995. An unmodified hetadecapeptide induces competence for genetic transformation in Streptococcus pneumoniae. Proc. Natl. Acad. Sci. USA 92, 11140-11144
  16. Havarstein, L.S., P. Gaustad, I.F. Nes, and D.A. Morrison. 1996. Identification of the streptococcal competence-pheromone receptor. Mol. Microbiol. 21, 863-869 https://doi.org/10.1046/j.1365-2958.1996.521416.x
  17. Lacks, S. and R.D. Hotchkiss. 1960. A study of the genetic material determining an enzyme in Pneumococcus. Biochim. Biophys. Acta. 22, 508–518
  18. Lee, M.S. and D.A. Morrison. 1999. Identification of a new regulator in Streptococcus pneumoniae linking quorum sensing to competence for genetic transformation. J. Bacteriol. 181, 5004- 5016
  19. Morrison, D.A. and M.F. Baker. 1979. Competence for genetic transformation in Pneumoncoccus depends on synthesis of a small set of proteins. Nature 282, 215-217 https://doi.org/10.1038/282215a0
  20. Mortier-Barriere, I., A. de Saizieu, J.P. Claverys, and B. Martin. 1998. Competence-specific induction of recA is required for full recombination proficiency during transformation in Streptococcus pneumoniae. Mol. Microbiol. 27, 159-170 https://doi.org/10.1046/j.1365-2958.1998.00668.x
  21. Paton, J.C., P.W. Andrew, G.J. Boulnois, and T.J. Mitchell. 1993. Molecular analysis of the pathogenicity of Streptococcus pneumoniae: the role of pneumococcal proteins. Annu. Rev. Microbiol. 47, 89-115 https://doi.org/10.1146/annurev.mi.47.100193.000513
  22. Pestova, E.V., L.S. Havarstein, and D.A. Morrison. 1996. Regulation of competence for genetic transformation in Streptococcus pneumoniae by an auto-induced peptide phermone and a twocomponent regulatory system. Mol. Microbiol. 21, 853-862 https://doi.org/10.1046/j.1365-2958.1996.501417.x
  23. Polissi, A., A. Pontiggia, G. Feger, M. Altieri, H. Mottl, L. Ferrari, and D. Simon. 1998. Large-scale identification of virulence genes from Streptococcus pneumoniae. Infect. Immun. 66, 5620-5629
  24. Poulsen, K., J. Reinholdt, C. jespersgaard, K. Boye, T.A. Brown, M. Hauge, and M. Kilian. 1998. A comprehensive genetic study of streptococcal immunoglobulin A1 proteases: evidence for recombination within and between species. Infect. Immun. 66, 181-190
  25. Pozzi, G., L. Masala, F. Iannell, R. Manganelli, L.S. Havarstein, L. Piccoli, D. Simon, and D.A. Morrison. 1996. Competence for genetic transformation in encapsulated strains of Streptococcus pneumoniae: two allelic variants of the peptide pheromone. J. Bacteriol. 178, 6087-6090 https://doi.org/10.1128/jb.178.20.6087-6090.1996
  26. Rosenow, C., P. Ryan, J.N. Weiser, S. Johnson, P. Fontan, A. Ortqvist, and H.R. Masure. 1997. Contribution of novel cholinebinding proteins to adherence, colonization and immunogenicity of Streptococcus pneumoniae. Mol. Microbiol. 25, 819-29 https://doi.org/10.1111/j.1365-2958.1997.mmi494.x
  27. Sampson, J.S., S.P. O'Connor, A.R, J.A. Tharpe, H. Russell. Stinson. 1994. Cloning and nucleotide sequence analysis of psaA, the Streptococcus pneumoniae gene encoding a 37-Kilodalton protein homologous to previously reported Streptococcus sp. adhesins. Infect. Inmmun. 62, 319-324
  28. Talkington, D.F., B.G. Brown, J.A. Tharpe, A. Koenig, and H. Russell. 1996. Protection of mice against fatal pneumococcal challenge by immenization with pnuemococcal surface adhesin A (PsaA). Microb. Pathog. 21, 17-22 https://doi.org/10.1006/mpat.1996.0038
  29. Tomasz, A. 1965. Control of the competent state on Pneumococcus by a hormone-like cell product: an example for a new type of regulatory mechanism in bacteria. Nature 208, 215-217
  30. Zwijnenburg, P.J., T. Van der Poll, S. Florquin, S.J. van Deventer, J.J. Roord, and A.M. van Furth. 2001. Experimental pneumococcal meningitis in mice: a model of intranasal infection. J. Infect. Dis. 183, 1143-1146 https://doi.org/10.1086/319271
  31. Wang, Y., Y.B. Yin, I. Shiro, J.Y. Luo, and X.M. Zhang. 2003. Investigation of competence-induced virulence of Streptococcus pneumoniae. Chinese journal of microbiology and immunology 23, 165-168