The hemagglutinating factor (hemagglutinin) of Bacteroides gingivalis was prepared from the supernatant of a 5-day diffusate broth culture by ammonium sulfate precipitation and column chromatography with a hydrophobic column of Phenyl-Sepharose CL-4B, DEAE-Sephadex A-50, and Sephadex G-100 gel filtration. The hemagglutinating activity of the preparation was 53.3 times higher than that of ammonium sulfate precipitate. In electron microphotographs, hemagglutinin appears to have a vesicle or tubelike structure. The hemagglutinating activity of intact cells was completely destroyed by heating at 100°C for 10 min, but the activity of extracted hemagglutinin was heat stable. The activity of hemagglutinin was inhibited by L-arginine and L-lysine and partially inhibited by phospholipase D, but it was not affected by proteolytic enzymes, neuraminidase, hyaluronidase, lipase, phospholipase A and C, or sugars. The B. gingivalis hemagglutinin appeared to be comprised mainly of a 40,000-molecular-weight material. The Fab fragment of immunoglobulin G prepared from rabbit antiserum to whole cells of B. gingivalis and monoclonal antibody against the hemagglutinin bound to the cell surface and inhibited the hemagglutinating activity of both the cells and the purffied hemagglutinin.

Serum and gingival crevicular fluid from normal healthy adults and patients with periodontitis were screened for immunoglobulin G antibodies to antigens from Bacteroides gingivalis 381, Bacteroides intermedius 24, Bacteroides loescheii ATCC 15930, Fusobacterium nucleatum ATCC 25586, Eikenella corrodens 1073, Actinobacillus actinomycetemcomitans ATCC 29522, and Capnocytophaga sp. strain M-12. Immunoglobulin G antibody titers to the antigens were measured by an enzyme-linked immunosorbent assay. The antibody levels to B. gingivalis in serum and gingival crevicular fluid were significantly higher in the samples from patients with periodontitis than in samples from healthy individuals. Although there were individual differences within patient groups, a positive correlation (P < 0.01) was found between the serum immunoglobulin G levels to B. gingivalis and the development of periodontitis. The antibodies to F. nucleatum (P < 0.05), E. corrodens (P < 0.05), and A. actinomycetemcomitans were slightly higher in patients with periodontitis than in normal subjects. There were no remarkable differences between the two groups in titers to B. intermedius, B. Ioescheii, and Capnocytophaga sp.

The initial event in colonization of the subgingival area by B. gingivalis is its attachment to host cells and Gram-positive bacteria in pre-formed plaque. The level of B. gingivalis is partly governed by products of other plaque bacteria, especially by sanguicin. Once B. gingivalis resides in its nidus and starts to proliferate, expulsion of pre-existing residents may occur, especially of attached Gram-positive bacteria, through the inhibitory action of the B. gingivalis product, hematin. The bacteriocin produced by black-pigmented Bacteroides also seems to play an important role in their establishment. Melaninogenicus possessed strong inhibitory activity against Actinomyces species. This was not completely confirmed with fresh isolates of B. gingivalis from advanced periodontitis patients. Various factors other than inhibitory substances produced by B. gingivalis and related bacteria can also affect the colonization of this species. Since the crevice area is influenced by gingival fluid, the nature of specific antibody and the other affecting components should be considered collectively with the interaction between new predominant colonizers and other pre-existing residents.

The relationship between the serum IgG antibody titer against seven species of Gram-negative periodontopathic bacteria and clinical parameters (including plaque index, gingival index, periodontal pocket depth, and alveolar bone loss) was studied in 38 subjects. IgG antibody titer against the sonicated antigens was determined by micro-ELISA. A statistically significant correlation was found between the serum antibody titer against B. gingivalis and the degree of clinical parameters, especially pocket depth. The serum IgG levels against the seven micro-organisms in 16 periodontal patients before and after clinical treatment were also determined. Responses to B. gingivalis decreased (p less than 0.001), whereas responses to E. corrodens (p less than 0.01) increased slightly. No marked differences were noted between pre- and post-treatment sera in titers against B. intermedius, B. loescheii, F. nucleatum, A. actinomycetemcomitans, and C. ochracea.

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