Human Gingival Epithelial Cells Produce Chemotactic Factors Interleukin‐8 and Monocyte Chemoattractant Protein‐1 After Stimulation With Porphyromonas gingivalis via Toll‐Like Receptor 2

• DOI: 10.1902/jop.2004.75.3.370
• Publication Date: 2005-02-02T12:52:16Z
• Authors: Yutaka Kusumoto; Hiroyuki Hirano; Keiko Saitoh; Satoru Yamada; Masahide Takedachi; Takenori Nozaki; Yasuhiro Ozawa; You Nakahira; Teruyuki Saho; Hideji Ogo; Yoshio Shimabukuro; Hiroshi Okada; Shinya Murakami
• Keywords: Lipopolysaccharides; Membrane Glycoproteins; Antigens, Polyomavirus Transforming; Interleukin-8; Gingiva; NF-kappa B; Epithelial Cells; Receptors, Cell Surface; Statistics, Nonparametric; Toll-Like Receptor 2; Toll-Like Receptor 4; Toll-Like Receptor 5; 03 medical and health sciences; 0302 clinical medicine; Fimbriae, Bacterial; Humans; RNA, Messenger; Coloring Agents; Porphyromonas gingivalis; Cells, Cultured; Chemokine CCL2; Signal Transduction

Background: The mechanism of stimulation of human gingival epithelial cells (HGEC) by Porphyromonas gingivalis (Pg) has not been fully clarified yet. In order to investigate the possible activation of HGEC by Pg through Toll‐like receptors (TLRs), we analyzed the production of chemotactic factors and the activated nuclear factor‐kappa B (NF‐κB).Methods: The mRNA expression of TLRs and the protein expression of TLR2 and TLR4 in HGEC and gingival tissue were assessed using reverse transcription‐polymerase chain reaction (RT‐PCR) assay and immunohistochemical staining. Primary cultured HGEC (nHGEC) and HGEC transformed by simian virus 40 T antigen (OBA‐9) were activated by a sonic extract (SE) of Pg to examine cytokine production and NF‐κB activation using enzyme‐linked immunosorbant assay (ELISA). In addition, Pg mediated activation of NF‐κB in a TLR2‐transfectant was also investigated.Results: RT‐PCR results revealed that HGEC expressed mRNA of TLR2, TLR4, TLR5, and TLR9, although the expression pro‐ files of each cell line were slightly different. In addition, immunostaining revealed the prominent expression of TLR2 not only in nHGEC, but also in the gingival epithelium of the tissue specimen. Interestingly, nHGEC and OBA‐9 secreted IL‐8 and monocyte chemoattractant protein (MCP)‐1 upon stimulation with Pg SE more efficiently than LPS and fimbriae of Pg. Furthermore, Pg SE increased the activated NF‐κB not only in OBA‐9, but also in 293T cells transfected with the human TLR2 gene.Conclusion: TLR2 participates, at least partly, in the signaling pathway to induce chemokine production in gingival epithelium as a reaction against Pg component(s), probably other than lipopolysaccharide and fimbriae. J Periodontol 2004;75:370‐379.