Immunobiological activities of a 55-kilodalton cell surface protein of Prevotella intermedia ATCC 25611

Abstract

A protein was extracted from whole cells of Prevoteila intermedia ATCC 25611 with sodium lauroylsarcosine and purified by chromatography on a DEAE-Sepharose fast-flow column. The apparent molecular weight of the protein was 55,000. A mouse polyclonal antibody specific for the protein recognized the cell surface structure of P. intermedia and also reacted with proteins in lysates of other black-pigmented anaerobic bacteria, such as Porphyromonas endodontalis and Prevotella melaninogenica, but not with those in lysates of Porphyromonas gingivalis or with the purified fimbriae of P. gingivalis 381. The N-terminal sequence of the 55-kDa protein showed only low homology with the cell surface proteins of any black-pigmented bacteria reported to date. The level of immunoglobulin G antibody to the antigen was higher in the sera of patients with periodontitis than in the sera of healthy volunteers. The protein induced interleukin-la,-1pI,-6, and-8 and tumor necrosis factor alpha in human peripheral blood mononuclear cell cultures and interleukin-li and-6 in human umbilical vascular endothelial cell and gingival fibroblast cultures. The protein induced interleukin-6 and tumor necrosis factor alpha activities in peritoneal macrophages from C3H/HeJ as well as from C3H/HeN mice and also induced cytokine activities in the sera of both strains of mice primed with muramyldipeptide.

Bacterial modulins: a novel class of virulence factors which cause host tissue pathology by inducing cytokine synthesis.

“…At 4 g/ml, fimbriae from this organism were able to induce IL-1␤ and GM-CSF gene expression in mouse embryonic calvarial bone cells (107). Matsushita et al (148) reported that a 55-kDa cell surface protein, possibly a fimbrial protein, from the periodontopathogenic organism Prevotella intermedia was able to stimulate the release of IL-1␣, IL-1␤, IL-6, and IL-8 from human PBMCs when it was present at a concentration of 0.1 g/ml but that TNF release required the fimbrial protein to be present at Ͼ1 g/ml. The potencies of the fimbriae were generally similar to those of LPS from the organism.…”

Conservation of fimbriae and the hemagglutinating adhesin HA-Ag2 among Porphyromonas gingivalis strains and other anaerobic bacteria studied by epitope mapping analysis

“…The anti-Pg-II fimbria MAbs described by Ogawa et al (23) do not recognize the 41-kDa subunit in P. gingivalis 381. Furthermore, no sequence homology was found between Pg-II fimbriae and the 50-kDa (33) and 53-kDa (15) proteins from P. gingivalis 381, the 51 kDa (12) and 67 kDa (15) proteins from P. gingivalis ATCC 33277, or the 55-kDa protein from Prevotella intermedia (20).…”

Lipopolysaccharide Enhances the Production of Vascular Endothelial Growth Factor by Human Pulp Cells in Culture

“…An upregulated production of VEGF in dental pulp may therefore enhance vascular permeability and the accumulation of inflammatory cells and increase the blood pressure, which results in the irreversible inflammation of dental pulp. However, VEGF also promotes the chemotaxis, proliferation, and differentiation of HPC (15,16). In the recovery phase of inflamed pulp tissue, a sufficient supply of blood and angiogenesis are required.…”

“…One of these components, lipopolysaccharide (LPS), is a potent inducer of pulpitis (37). LPS induces many inflammatory cytokines such as interleukin-1 (IL-1), IL-6, IL-8, and tumor necrosis factor alpha (TNF-␣) from human peripheral blood mononuclear cells and gingival fibroblasts (16,28). LPS also induces increased arachidonic acid metabolism in the pulp; LPS was shown to induce PGE 2 and PGI 2 production, resulting in an increase of vascular permeability and neutrophil infiltration (18).…”