In this study, evidence is presented that mixed infection with the periodontal pathogens Porphyromonas gingivalis and Actinobacillus actinomycetemcomitans results in a synergistic effect in their pathogenicity and in their ability to induce humoral and cellular host responses. BALB/c mice were injected subcutaneously on the back with P. gingivalis ATCC 53977, A. actinomycetemocomitans 75 or a mixture of both bacteria. Samples of blood and fluid from abscesses formed at the site of injection (first degree) or distant from the injection site were collected for microbiologic analysis. Serum and spleens were obtained for evaluation of humoral and cellular responses to P. gingivalis and A actinomycetemocomitans. Mice injected with A. actinomycetemcomitans had first-degree lesions only, whereas mice injected with P. gingivalis and A. actinomycetemcomitans had lesions at first- and second-degree sites from which both bacterial species were isolated. A serum anti-P. gingivalis response was induced in P. gingivalis-injected mice, which was higher in mice injected with P. gingivalis and A. actinomycetemcomitans. This pattern was not seen in the anti-A, actinomycetemcomitans response. Lymphoproliferative responses to phytohemagglutinin, Escherichia coli lipopolysaccharide and P. gingivalis of spleen cells from infected mice were decreased, especially following co-infection. Furthermore, co-infection of mice resulted in the greatest decrease in the number of CD5+, especially CD4+ lymphocytes.

To better understand the role of the capsular polysaccharide in the virulence of Porphyromonas gingivalis, the effect of immunization with a polysaccharide-protein conjugate on experimental murine infection was evaluated. The conjugate was prepared using polysaccharide isolated from P. gingivalis strain ATCC 53977 and bovine serum albumin. One group of 22 mice was immunized by intraperitoneal injection with the conjugate and a control group of 25 mice was similarly immunized with bovine serum albumin. Serum antibody reactive to the polysaccharide, as determined by enzyme-linked immunosorbent assay, was elevated in the group of mice immunized with the polysaccharide-protein conjugate but not in the mice immunized with bovine serum albumin. Both groups of mice were challenged with P. gingivalis strain ATCC 53977 (10(10) cells) administered subcutaneously on the dorsal surface. Following challenge, the mice immunized with the polysaccharide-protein conjugate appeared healthier and demonstrated less weight loss than did the control group of mice. Ulcerative lesions at secondary locations were smaller in mice immunized with the polysaccharide-protein conjugate. Thus, immunization of mice with a conjugate containing P. gingivalis polysaccharide could reduce the severity of but not prevent an invasive infection with P. gingivalis.