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Updated: Oct 21, 2025
Porphyromonas gingivalis as a Model Organism for Assessing Interaction of Anaerobic Bacteria with Host Cells
Published on: December 17, 2015
Generation and functional characterization of recombinant Porphyromonas gingivalis W83 FimA
S Groeger1, M Hudel2, S Zechel2
1Department of Periodontology, Justus-Liebig-University of Giessen, Germany.
Abstract:
Porphyromonas gingivalis (P. gingivalis) is regarded as a keystone pathogen in destructive periodontal diseases. It expresses a variety of virulence factors, amongst them fimbriae that are involved in colonization, invasion, establishment and persistence of the bacteria inside the host cells. The fimbriae also were demonstrated to affect the host immune-response mechanisms. The major fimbriae are able to bind specifically to different host cells, amongst them peripheral blood monocytes. The interaction of these cells with fimbriae induces release of cytokines such as interleukin-1 (IL-1), IL-6, and tumor necrosis factor-α (TNF-α). The aim of this study was to generate recombinant major FimA protein from P. gingivalis W83 fimbriae and to prove its biological activity. FimA of P. gingivalis W83 was amplified from chromosomal DNA, cloned in a vector and transferred into Listeria innocua. (L. innocua).The expressed protein was harvested and purified using FPLC via a His trap HP column. The identity and purity was demonstrated by gel-electrophoresis and mass-spectrometry. The biological activity was assessed by stimulation of human oral epithelial cells and peripheral blood monocytes with the protein and afterwards cytokines in the supernatants were quantified by enzyme linked immunosorbent assay (ELISA) and cytometric bead array. Recombinant FimA could successfully be generated and purified. Gel-electrophoresis and mass-spectrometry confirmed that the detected sequences are identical with FimA. Stimulation of human monocytes induced the release of high concentrations of IL-1β, IL-6, IL-10 and TNF-α by these cells. In conclusion, a recombinant FimA protein was established and its biological activity was proven. This protein may serve as a promising agent for further investigation of its role in periodontitis and possible new therapeutic approaches.
Insights
Porphyromonas gingivalis fimbriae protein (FimA) was successfully produced and purified. This recombinant FimA triggers immune responses, releasing cytokines from human monocytes, suggesting its role in periodontitis.
Area of Science:
- Microbiology
- Immunology
- Periodontology
Background:
- Porphyromonas gingivalis is a key pathogen in periodontal disease.
- Bacterial fimbriae are crucial virulence factors involved in host cell colonization and immune modulation.
- Fimbriae binding to host cells, like monocytes, stimulates cytokine release.
Purpose of the Study:
- To generate recombinant major FimA protein from P. gingivalis W83.
- To confirm the biological activity of the recombinant FimA protein.
Main Methods:
- FimA gene amplification, cloning into Listeria innocua, and protein expression.
- Protein purification using FPLC and His trap HP column.
- Confirmation of protein identity and purity via gel electrophoresis and mass spectrometry.
- Assessment of biological activity by stimulating human monocytes and quantifying cytokine release using ELISA and cytometric bead array.
Main Results:
- Recombinant FimA protein was successfully generated and purified.
- Gel electrophoresis and mass spectrometry confirmed FimA identity and purity.
- Stimulation of human monocytes with recombinant FimA induced significant release of IL-1β, IL-6, IL-10, and TNF-α.
Conclusions:
- A biologically active recombinant FimA protein from P. gingivalis W83 was successfully produced.
- The recombinant FimA protein effectively stimulates cytokine release from human monocytes.
- This protein is a valuable tool for studying P. gingivalis's role in periodontitis and developing new therapies.
