Inhibition of experimental periodontitis by a monoclonal antibody against Porphyromonas gingivalis HA2

It is known that complexes of the multi-protein, gingipain, possess heme binding domains (hemagglutinin 2, HA2) that bind hemoglobin to provide heme and iron to the bacterium, Porphyromonas gingivalis. The DHYAV-MISK peptide sequence was proposed to act as an inhibitor of hemin binding, and thus, it might be used to control or prevent periodontal disease. In this study, we created a monoclonal antibody (mAb) that targeted the DHYAVMISK peptide, aimed to determine whether it could inhibit the growth of P. gingivalis in vitro, and block its induction of experimental periodontitis and subsequent bone loss. Peptide DGFPG-DHYAVMISK conjugated to KLH (DK-KLH) was synthetic, and injected subcutaneously into BALB/c mice to generate specific mAbs with the hybridoma technique. We isolated mAb 1H11, which showed specific binding to DK. When we incubated these mAbs with P. gingivalis in vitro for 18 h, bacterial growth was significantly lower in cultures treated with mAb 1H11 compared to those treated with control (PBS; P < 0.05). Next, we induced experimental periodontitis in mouse models with a silk ligature and a P. gingivalis infection. When we injected the mAbs into the gingival sulcus, the group treated with mAb 1H11 displayed a reduction in bone loss compared to the other treatment groups. Thus, mAb 1H11 might provide protection against a P. gingivalis infection. Accordingly, this antibody could serve as a candidate therapy for periodontitis or other infections caused by P. gingivalis.