CHARACTERIZATION OF IMMUNOGLOBULIN-G-DEGRADING PROTEASES OF PREVOTELLA-INTERMEDIA AND PREVOTELLA-NIGRESCENS

Degradation of immunoglobulins is thought to be an important factor in the causation of periodontal diseases by hindering local host defenses and by providing nutrients to the periodontal microflora, In this study, we characterized the proteolytic activity against human immunoglobulin G (IgG) of 20 strains of Prevotella intermedia and Prevotella nigrescens isolated from periodontal pockets and oral abscesses. IgG degradation was studied by sodium dodecyl sulphate-polyacrylamide gel electrophoresis. All strains degraded IgG within 48 h after growth in trypticase-yeast extract medium (TY) supplemented with 0.3% IgG, Incorporating IgG in TY broth enhanced bacterial growth. Protease profiles (zymography), which revealed the presence of 1-4 IgG-degrading proteolytic bands in bacterial cell extracts, became more complex after growth in the presence of IgG, A 38-kDa protease capable of degrading IgG nonspecifically was present in almost all strains. The proteolytic activity was mainly located on the surface of the cell envelope. Two strains of P. intermedia and P. nigrescens ATCC 33563 were selected for further studies. Bacterial cell suspensions in phosphate-buffered saline completely degraded human IgG, IgA and IgM within 24 h. This activity depended on reducing conditions and was inhibited at temperatures above 50 degrees C. The pH optimum of immunoglobulin degradation was at pH 7. Strains cultured at 42 degrees C showed a markedly reduced capacity to degrade IgG. Inhibition studies revealed that breakdown of IgG was caused by a cysteine protease(s). The capacity of P. intermedia and P. nigrescens to degrade immunoglobulins may explain their association with polymicrobial oral diseases.