This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by O'Brien-Simpson, N. M.
Right arrow Articles by Reynolds, E. C.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by O'Brien-Simpson, N. M.
Right arrow Articles by Reynolds, E. C.

 Previous Article  |  Next Article 

Infection and Immunity, July 2000, p. 4055-4063, Vol. 68, No. 7
0019-9567/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

RgpA-Kgp Peptide-Based Immunogens Provide Protection against Porphyromonas gingivalis Challenge in a Murine Lesion Model

Neil M. O'Brien-Simpson, Rita A. Paolini, and Eric C. Reynolds*

Oral Health Sciences Unit, School of Dental Science, The University of Melbourne, Melbourne, Victoria 3000, Australia

Received 22 November 1999/Returned for modification 1 March 2000/Accepted 11 April 2000

Porphyromonas gingivalis, a gram-negative bacterium, has been linked to the onset and progression of periodontitis, a chronic inflammatory disease of the supporting tissues of the teeth. A major virulence factor of P. gingivalis is an extracellular complex of Arg- and Lys-specific proteinases and adhesins designated the RgpA-Kgp complex (formerly the PrtR-PrtK complex). In this study we show that the RgpA-Kgp complex, when used as an immunogen with incomplete Freund adjuvant (IFA), protects against challenge with invasive and noninvasive strains of P. gingivalis in the murine lesion model. We identified a variety of peptide vaccine candidates from the RgpA and Kgp polyprotein sequences that involved the putative active site histidine of both proteinases and five repeat motifs in the adhesin domains of both polyproteins implicated in aggregation and binding to host substrates, designated adhesin-binding motif (ABM) peptides. These peptides were synthesized using standard, solid-phase protocols for 9-fluorenylmethoxy carbonyl chemistry with S-acetylmercaptoacetic acid (SAMA) as the N-terminal residue. The SAMA-peptides were then conjugated to diphtheria toxoid and used with IFA to immunize BALB/c mice. Both active-site peptides and three of the five ABM peptides gave protection (P < 0.005) against challenge with P. gingivalis in the murine lesion model. The three ABM peptide sequences that conferred protection exist within a 100-residue span in the RgpA44 and Kgp39 adhesins of the RgpA-Kgp complex. Protective anti-RgpA-Kgp complex mouse antisera recognized the RgpA27, Kgp39, and RgpA44 adhesins in an immunoblot. Epitope mapping of the RgpA27 adhesin using the protective anti-RgpA-Kgp antisera identified a major protective epitope that mapped immediately N terminal to one of the protective ABM peptides in the 100-residue span in RgpA44 and Kgp39. This identified protective epitope contains clusters of basic residues spatially surrounded by hydrophobic amino acids, a finding which is characteristic of a heparin binding motif.

* Corresponding author. Mailing address: Oral Health Sciences Unit, 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}dent.unimelb.edu.au.

Infection and Immunity, July 2000, p. 4055-4063, Vol. 68, No. 7
0019-9567/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Tam, V., O'Brien-Simpson, N. M., Pathirana, R. D., Frazer, L. T., Reynolds, E. C. (2008). Characterization of T Cell Responses to the RgpA-Kgp Proteinase-Adhesin Complexes of Porphyromonas gingivalis in BALB/c Mice. J. Immunol. 181: 4150-4158 [Abstract] [Full Text]  
  • Pathirana, R. D., O'Brien-Simpson, N. M., Veith, P. D., Riley, P. F., Reynolds, E. C. (2006). Characterization of proteinase-adhesin complexes of Porphyromonas gingivalis.. Microbiology 152: 2381-2394 [Abstract] [Full Text]  
  • Teng, Y.-T.A. (2006). Protective and Destructive Immunity in the Periodontium: Part 2--T-cell-mediated Immunity in the Periodontium. JDR 85: 209-219 [Abstract] [Full Text]  
  • O'Brien-Simpson, N. M., Pathirana, R. D., Paolini, R. A., Chen, Y.-Y., Veith, P. D., Tam, V., Ally, N., Pike, R. N., Reynolds, E. C. (2005). An Immune Response Directed to Proteinase and Adhesin Functional Epitopes Protects against Porphyromonas gingivalis-Induced Periodontal Bone Loss. J. Immunol. 175: 3980-3989 [Abstract] [Full Text]  
  • Gibson, F. C. III, Gonzalez, D. A., Wong, J., Genco, C. A. (2004). Porphyromonas gingivalis-Specific Immunoglobulin G Prevents P. gingivalis-Elicited Oral Bone Loss in a Murine Model. Infect. Immun. 72: 2408-2411 [Abstract] [Full Text]  
  • Chen, Y.-Y., Cross, K. J., Paolini, R. A., Fielding, J. E., Slakeski, N., Reynolds, E. C. (2002). CPG70 Is a Novel Basic Metallocarboxypeptidase with C-terminal Polycystic Kidney Disease Domains from Porphyromonas gingivalis. J. Biol. Chem. 277: 23433-23440 [Abstract] [Full Text]  
  • Rajapakse, P. S., O'Brien-Simpson, N. M., Slakeski, N., Hoffmann, B., Reynolds, E. C. (2002). Immunization with the RgpA-Kgp Proteinase-Adhesin Complexes of Porphyromonas gingivalis Protects against Periodontal Bone Loss in the Rat Periodontitis Model. Infect. Immun. 70: 2480-2486 [Abstract] [Full Text]  
  • O'Brien-Simpson, N. M., Paolini, R. A., Hoffmann, B., Slakeski, N., Dashper, S. G., Reynolds, E. C. (2001). Role of RgpA, RgpB, and Kgp Proteinases in Virulence of Porphyromonas gingivalis W50 in a Murine Lesion Model. Infect. Immun. 69: 7527-7534 [Abstract] [Full Text]  
  • Gibson, F. C. III, Genco, C. A. (2001). Prevention of Porphyromonas gingivalis-Induced Oral Bone Loss following Immunization with Gingipain R1. Infect. Immun. 69: 7959-7963 [Abstract] [Full Text]  
  • Yonezawa, H., Ishihara, K., Okuda, K. (2001). Arg-Gingipain A DNA Vaccine Induces Protective Immunity against Infection by Porphyromonas gingivalis in a Murine Model. Infect. Immun. 69: 2858-2864 [Abstract] [Full Text]  
  • Kuboniwa, M., Amano, A., Shizukuishi, S., Nakagawa, I., Hamada, S. (2001). Specific Antibodies to Porphyromonas gingivalis Lys-Gingipain by DNA Vaccination Inhibit Bacterial Binding to Hemoglobin and Protect Mice from Infection. Infect. Immun. 69: 2972-2979 [Abstract] [Full Text]  
  • Chen, T., Nakayama, K., Belliveau, L., Duncan, M. J. (2001). Porphyromonas gingivalis Gingipains and Adhesion to Epithelial Cells. Infect. Immun. 69: 3048-3056 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»