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Porphyromonas gingivalis and Treponema denticola are major pathogens of periodontal disease. Coaggregation between microorganisms plays a key role in the colonization of the gingival crevice and the organization of periodontopathic biofilms. We investigated the involvement of surface ligands of P. gingivalis in coaggregation. Two triple mutants of P. gingivalis lacking Arg-gingipain A (RgpA), Lys-gingipain (Kgp) and Hemagglutinin A (HagA) or RgpA, Arg-gingipain B (RgpB) and Kgp showed significantly decreased coaggregation with T. denticola, whereas coaggregation with a major fimbriae (FimA)-deficient mutant was the same as that with the P. gingivalis wild-type parent strain. rgpA, kgp and hagA code for proteins that contain 44 kDa Hgp44 adhesin domains. The coaggregation activity of an rgpA kgp mutant was significantly higher than that of the rgpA kgp hagA mutant. Furthermore, anti-Hgp44 immunoglobulin G reduced coaggregation between P. gingivalis wild type and T. denticola. Treponema denticola sonicates adhered to recombinant Rgp domains. Coaggregation following co-culture of the rgpA kgp hagA mutant expressing the RgpB protease with the rgpA rgpB kgp mutant expressing the unprocessed HagA protein was enhanced compared with that of each triple mutant with T. denticola. These results indicate that the processed P. gingivalis surface Hgp44 domains are key adhesion factors for coaggregation with T. denticola.

The aim of this investigation was to analyze the effects of early life exposure to periodontopathic bacterial lipopolysaccharides on immunoresponse. Newborn BALB/c mice were subcutaneously injected with 20 ng lipopolysaccharide of Porphyromonas gingivalis, Actinobacillus actinomycetemcomitans or Escherichia coli daily for 2 days, starting within 24 h after birth. The treated mice were given intraperitoneal injections of bovine serum albumin at 180 and 187 days of age. Seventeen hours after each injection, the mice were bled and sera were separated. Their sera were tested in an enzyme-linked immunosorbent assay system. The mean interleukin-4, interleukin-5, interleukin-6 and immunoglobulin E levels in the sera of mice treated neonatally with P. gingivalis lipopolysaccharide were significantly higher than those of the controls. However, in all cases, no significant difference was noted between mice treated neonatally with A. actinomycetemcomitans- or E. coli lipopolysaccharide and control mice. These data suggest that neonatal exposure to P. gingivalis lipopolysaccharide induces changes in immunological responses when the mice reach maturity.

We studied the effect of antibodies against Porphyromonas gingivalis gingipain domains, preparing them against three recombinant fragments of RgpA (catalytic domain, r-Rgp CAT; hemagglutinin domains, r-Rgp 44 and r-Rgps 15-27) and one fragment of Kgp (catalytic domain, r-Kgp CAT). Enhancement of opsonization and killing by human polymorphonuclear leukocytes were measured in the noninvasive FDC 381 and invasive W50 strains of P. gingivalis. Anti-r-Rgp 44 was the most effective in both strains of P. gingivalis. The present findings lead us to recommend RgpA 44 as a candidate immunogen for vaccines against P. gingivalis.

A protease of Treponema denticola, dentilisin, is thought to be part of a complex with 43- and 38-kDa proteins. A sequence encoding a 43-kDa protein was located in the 3' region of the prcA gene upstream of the dentilisin gene (prtP). The 43-kDa protein was apparently generated from digestion of PrcA. To clarify the function of the protein, we constructed a mutant of the 43-kDa protein following homologous recombination. The mutant lacked detectable dentilisin activity. Immunoblot analysis demonstrated that the dentilisin protein was degraded in the mutant. The results of real-time polymerase chain reaction suggested that prtP mRNA expression in the mutant was somewhat decreased compared with the wild-type strain. These data suggest that the 43-kDa protein is involved in the stabilization of the dentilisin protein.

Periodontopathic Campylobacter rectus strains possess 41- and 68-kDa proteinaceous antigens which share antigenicity with antigens of Helicobacter pylori strains. H. pylori strains have a 54-kDa antigen which reacts with C. rectus strains. We found that the salivary IgA levels against H. pylori were correlated with those against C. rectus. These cross-reactive antigens of C. rectus may affect the serological diagnosis of H. pylori infections, especially when saliva is used. It is possible that these cross-reacting antigens may relate to the induction of immunopathological responses against both microorganisms.