|Budget Amount *help
¥2,900,000 (Direct Cost : ¥2,900,000)
Fiscal Year 2003 : ¥1,100,000 (Direct Cost : ¥1,100,000)
Fiscal Year 2002 : ¥1,800,000 (Direct Cost : ¥1,800,000)
The aim of this study is to investigate the response of oral mucous epithelial cells to infection of a major bacterial pathogens of periodontalis, Porphyromonas gin givalis. Since lipopolysaccharide (LPS) is one of major vilulence factors of P. gin givalis, effects of P. gingivalis LPS on mouse sulcular epithelium cell line, GEL cells, were studies. The obtained results are as follows:
1. Membrane receptors for LPS. P. gin givalis LPS enhanced accumulation of Toll-like receptor (TLR) 4 mRNA in GEl cells, but did not significantly affect on expression of CDL4 and TLR2 genes. These receptors were detectable on cytoplasmic membrane of GEl by analysis with a flowcytometer and immunohistochemistry by using specific antibodies. The expression level of CD14 in GEl cells was comparable to that in macrophage-like cell, J774. 1, whereas the levels of TLR2 and TLR4 were apparently lower.
2. Responses of suiclar epithelium against LPS. Cell proliferation of GEl increased by P. gin givalis LPS. Gene expression of inflammatory cytokines, such as TNFcz, IL-1β and IL-6, and chemokines, such as IL-8 and inflammatory protein 2, altered by incubation with P. gin givalis LPS. Concomitant with these responses, phosphorylation of Thr and Tyr residues of GE1 proteins was demonstrated by 2D-FAGE and ECL Western blotting.
These results suggested that sulclar epithelium cells could response to periodontal virulence factors of P.gin givalis LPS and that receptors for LPS may be different from those reported on macrophages.