Immunoglobulin G (IgG) class, but Not IgA or IgM, antibodies to peptides of the Porphyromonas gingivalis chaperone HtpG predict health in subjects with periodontitis by a fluorescence enzyme-linked immunosorbent assay

Abstract

Chaperones are molecules found in all cells and are critical in stabilization of synthesized proteins, in repair/removal of defective proteins, and as immunodominant antigens in innate and adaptive immunity. Subjects with gingivitis colonized by the oral pathogen Porphyromonas gingivalis previously demonstrated levels of anti-human chaperone Hsp90 that were highest in individuals with the best oral health. We hypothesized that similar antibodies to pathogen chaperones might be protective in periodontitis. This study examined the relationship between antibodies to P. gingivalis HtpG and clinical statuses of healthy and periodontitis-susceptible subjects. We measured the humoral responses (immunoglobulin G [IgG], IgA, and IgM) to peptides of a unique insert (P18) found in Bacteroidaceae HtpG by using a high-throughput, quantitative fluorescence enzyme-linked immunosorbent assay. Indeed, higher levels of IgG class anti-P. gingivalis HtpG P18 peptide (P < 0.05) and P18alpha, consisting of the N-terminal 16 amino acids of P18 (P < 0.05), were associated with better oral health; these results were opposite of those found with anti-P. gingivalis whole-cell antibodies and levels of the bacterium in the subgingival biofilm. When we examined the same sera for IgA and IgM class antibodies, we found no significant relationship to subject clinical status. The relationship between anti-P18 levels and clinical populations and individual subjects was found to be improved when we normalized the anti-P18alpha values to those for anti-P18gamma (the central 16 amino acids of P18). That same ratio correlated with the improvement in tissue attachment gain after treatment (P < 0.05). We suggest that anti-P. gingivalis HtpG P18alpha antibodies are protective in periodontal disease and may have prognostic value for guidance of individual patient treatment.

Figures

FIG. 1.

Normalization of anti-P. gingivalis levels to those for internal peptides. Values for anti-P18 peptide antibodies to the three subpeptides of P18 were normalized by dividing the concentration of each antisubpeptide antibody by the same values for the other individual subpeptides: anti-P18α/anti-P18γ (clear bars), anti-P18α/anti-P18β (hatched bars), and anti-P18β/anti-P18γ (filled bars). Means for all subjects were calculated and those for healthy and periodontitis-susceptible subjects compared by ANOVA (boxed insert). Point, mean; box, mean ± standard error; diamond, mean ± 1.96 × standard error.

FIG. 2.

P18α/P18γ antisubpeptide ratio correlates with clinical measurements at the baseline. Pearson R correlations were calculated for PD and CAL by using a linear fit for either PD (filled circles) or CAL (open squares) to scatter plots. The associated correlation coefficients (r values) and probabilities (P values) are displayed in the boxed insert.

FIG. 3.

Correlation of anti-P18α/anti-P18γ ratio with tissue improvement after periodontitis treatment. Pearson R correlations between percents reduction of PD (filled circle) and recovery of CAL (filled squares) (6 months after treatment) and anti-P18α/anti-P18γ ratio are shown. The associated correlation coefficients (r values) and probabilities (P values) are displayed in the boxed insert.