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Infection and Immunity, May 2000, p. 2704-2712, Vol. 68, No. 5
0019-9567/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Serum Immunoglobulin G (IgG) and IgG Subclass Responses to the RgpA-Kgp Proteinase-Adhesin Complex of Porphyromonas gingivalis in Adult Periodontitis

Neil M. O'Brien-Simpson, Catherine L. Black, Peter S. Bhogal, Steven M. Cleal, Nada Slakeski, Thomas J. Higgins, and Eric C. Reynolds*

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

Received 7 May 1999/Returned for modification 7 July 1999/Accepted 16 February 2000

Serum immunoglobulin G (IgG), IgM, and IgG subclass responses to the RgpA-Kgp proteinase-adhesin complex of Porphyromonas gingivalis were examined by enzyme-linked immunosorbent assay using adult periodontitis patients and age- and sex-matched controls. Twenty-five sera from subjects with adult periodontitis (diseased group) and 25 sera from healthy subjects (control group) were used for the study. Sera and subgingival plaque samples from 10 sites were collected from each patient at the time of clinical examination. The level of P. gingivalis in the plaque samples was determined using a DNA probe. Highly significant positive associations between the percentage of sites positive for P. gingivalis and measures of disease severity (mean pocket depth, mean attachment loss, and percentage of sites that bled on probing) were found. The diseased group had significantly higher specific IgG responses to the RgpA-Kgp complex than did the control group, and the responses were significantly associated with mean probing depths and percentage of sites positive for P. gingivalis. Analysis of the IgG subclass responses to the RgpA-Kgp complex revealed that the subclass distribution for both the diseased and control groups was IgG4 > IgG2 > IgG3 = IgG1. The IgG2 response to the complex was positively correlated with mean probing depth, whereas the IgG4 response was negatively correlated with this measure of disease severity. Immunoblot analysis of the RgpA-Kgp complex showed that sera from healthy subjects and those with low levels of disease, with high IgG4 and low IgG2 responses, reacted with the RgpA27, Kgp39, and RgpA44 adhesins; however, sera from diseased subjects with low IgG4 and high IgG2 responses reacted only with the RgpA44 and/or Kgp44 adhesins. Epitope mapping of the RgpA27 adhesin localized a major epitope recognized by IgG4 antibodies in sera from subjects with high IgG4 and low IgG2 responses to the RgpA-Kgp complex which was not recognized by sera from diseased subjects with low IgG4 and high IgG2 responses.

* 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, May 2000, p. 2704-2712, Vol. 68, No. 5
0019-9567/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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