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Porphyromonas gingivalis has been implicated as an important pathogen in the development of adult periodontitis, and its colonization of subgingival sites is critical in the pathogenic process. One potential virulence factor, hemagglutinin, may mediate bacteria attachment onto and penetration into host cells, as well as agglutinate and lyses erythrocytes to intake heme, an absolute requirement for growth. Toward the development of passive immunotherapy, the construction of a human type monoclonal antibody, which is capable of inhibiting the hemagglutinating ability, will be significant and important. The human mAbs, both exhibiting a high degree of specificity and affinity against the recombinant 130 kDa hemagglutinin domain protein have been prepared using XenoMouse technology. The constructed Xeno-mAbs, IgG2 subclass, significantly inhibited hemagglutination of P. gingivalis and its vesicles. The newly constructed Xeno-mAbs may prove to be useful for the development of passive immunization against periodontal diseases caused by P. gingivalis infection, pending the results of fertility study in disease mode.

Background: Porphyromonas gingivalis has been implicated as an important pathogen in the development of chronic periodontitis, and its colonization of subgingival sites is critical in the pathogenic process. One potential virulence factor, hemagglutinin, may mediate bacteria attachment onto and penetration into host cells, as well as agglutinate and lyse erythrocytes to intake heme, an absolute requirement for growth. We previously cloned the gene encoding the 130 kDa hemagglutinin domain (130k HMGD) and identified its functional domain. The construction of a human monoclonal antibody that is capable of inhibiting the hemagglutinating ability is significant and important toward the development of passive immunotherapy., Methods: Human lymphocytes isolated from a donor, who had high antibody titer against the recombinant 130k HMGD (r130k HMGD), were immortalized by Epstein-Barr virus, and specific antibody-producing B cells were established by panning using the r130k HMGD., Results: The constructed HuMAb-HMGD1, IgG subclass, recognized the r130k HMGD as well as the 43 and 49 kDa major bands in P. gingivalis cells and vesicles. The HuMAb-HMGD1 significantly inhibited hemagglutinating activity of P. gingivalis vesicles in a dose-dependent manner. Furthermore, the HuMAb-HMGD1 recognized the synthetic peptide, EGSNEFAPVQNLTGSSVG, which contains the functional domain of 130k HMGD., Conclusion: The newly constructed HuMAb-HMGD1 may prove to be useful for the development of passive immunization against periodontal diseases caused by P. gingivalis infection, pending the results of fertility study in disease mode.

Porphyromonas gingivalis has been implicated as an important pathogen in the development of adult periodontitis. This bacterium possesses hemagglutinating and hemolytic activities to attach and lyse erythrocytes. Hemolysis by this oral pathogen functions to provide heme-containing molecules for growth in the periodontal pocket. We previously constructed a monoclonal antibody using P. gingivalis vesicles as the immunogen, designated as MAb-Pg-vc, which inhibited vesicle-associated hemagglutinating activity. Furthermore, we cloned the gene encoding 130-kDa hemagglutinin (130-kDa HAG) and identified its functional motif for attachment to erythrocytes. Generally, bacterial cell attachment to erythrocytes is an important initial step for expressing hemolysis activity. In the present study, we examined the effect of MAb-Pg-vc on the hemolytic activity of P. gingivalis cells. The MAb-Pg-vc significantly inhibited the hemolytic activity and, further, this inhibitory activity was reduced by the synthetic peptide of the 130-kDa HAG functional motif.