Journalartikel

Jahr der Veröffentlichung: 2021

Seiten: 22-29

Zeitschrift: Journal of Biotechnology

Bandnummer: 340

ISSN: 0168-1656

eISSN: 1873-4863

DOI Link: https://doi.org/10.1016/j.jbiotec.2021.08.009

Verlag: Elsevier

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 cy-tokines such as interleukin-1 (IL-1), IL-6, and tumor necrosis factor-alpha (TNF-alpha). 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-1 beta, IL-6, IL-10 and TNF-alpha by these cells. In conclu-sion, 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.

Harvard-Zitierstil: Groeger, S., Hudel, M., Zechel, S., Chakraborty, T., Lochnit, G., Meyle, J., et al. (2021) Generation and functional characterization of recombinant Porphyromonas gingivalis W83 FimA, Journal of Biotechnology, 340, pp. 22-29. https://doi.org/10.1016/j.jbiotec.2021.08.009

APA-Zitierstil: Groeger, S., Hudel, M., Zechel, S., Chakraborty, T., Lochnit, G., Meyle, J., & Domann, E. (2021). Generation and functional characterization of recombinant Porphyromonas gingivalis W83 FimA. Journal of Biotechnology. 340, 22-29. https://doi.org/10.1016/j.jbiotec.2021.08.009