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Infection and Immunity, May 2002, p. 2480-2486, Vol. 70, No. 5
0019-9567/02/$04.00+0     DOI: 10.1128/IAI.70.5.2480-2486.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Immunization with the RgpA-Kgp Proteinase-Adhesin Complexes of Porphyromonas gingivalis Protects against Periodontal Bone Loss in the Rat Periodontitis Model

P. Sunethra Rajapakse, Neil M. O'Brien-Simpson, Nada Slakeski, Brigitte Hoffmann, and Eric C. Reynolds*

School of Dental Science, The University of Melbourne, Melbourne, Australia

Received 21 September 2001/ Returned for modification 18 December 2001/ Accepted 11 February 2002

A major virulence factor of Porphyromonas gingivalis is the extracellular noncovalently associated complexes of Arg-X- and Lys-X-specific cysteine proteinases and adhesins designated the RgpA-Kgp complexes. In this study we investigated the ability of RgpA-Kgp as an immunogen to protect against P. gingivalis-induced periodontal bone loss in the rat. Specific-pathogen-free Sprague-Dawley rats were immunized with either formalin-killed whole P. gingivalis ATCC 33277 cells with incomplete Freund's adjuvant, RgpA-Kgp with incomplete Freund's adjuvant, or incomplete Freund's adjuvant alone. The animals were then challenged by oral inoculation with live P. gingivalis ATCC 33277 cells. Marked periodontal bone loss was observed in animals immunized with incomplete Freund's adjuvant alone; this bone loss was significantly (P < 0.05) greater than that detected in animals immunized with formalin-killed whole cells or RgpA-Kgp or in unchallenged animals. There was no significant difference in periodontal bone loss between animals immunized with formalin-killed whole cells and those immunized with RgpA-Kgp. The bone loss in these animals was also not significantly different from that in unchallenged animals. DNA probe analysis of subgingival plaque samples showed that 100% of the animals immunized with incomplete Freund's adjuvant alone and challenged with P. gingivalis ATCC 33277 were positive for the bacterium. However, P. gingivalis ATCC 33277 could not be detected in subgingival plaque samples from animals immunized with formalin-killed whole cells or with RgpA-Kgp. Immunization with formalin-killed whole cells or RgpA-Kgp induced a high-titer serum immunoglobulin G2a response. Western blot analysis of RgpA-Kgp using pooled protective antisera taken from rats immunized with RgpA-Kgp revealed immunodominant bands at 44, 39, and 27 kDa. In conclusion, immunization with RgpA-Kgp restricted colonization by P. gingivalis and periodontal bone loss in the rat.

* Corresponding author. Mailing address: School of Dental Science, The University of Melbourne, 711 Elizabeth St., Melbourne, Victoria 3000, Australia. Phone: 61 3 9341 0270. Fax: 61 3 9341 0236. E-mail: e.reynolds{at}unimelb.edu.au.

Editor: R. N. Moore

Infection and Immunity, May 2002, p. 2480-2486, Vol. 70, No. 5
0019-9567/02/$04.00+0     DOI: 10.1128/IAI.70.5.2480-2486.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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