| Budget Amount *help |
¥3,400,000 (Direct Cost: ¥3,400,000)
Fiscal Year 2001: ¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 2000: ¥2,200,000 (Direct Cost: ¥2,200,000)
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| Research Abstract |
The aim of this study is to investigate the response of gingival epithelial cells to virulence factors from periodontal pathogens, Porphyromonas gingivalis, at the early onset of bacterial infection, using the mouse gingival epithelial cell line (GE cells) which we have established. The obtained results are as follows :
1. Immunohistochemical study with a confocal laser microscopy revealed existence of receptors for bacterial virulence factors, i.e. CDI4, Toll-like receptor (TLR)-2, and TLR-4, on the surface of unstimulated GE1 cells. This result suggested that gingival epithelial cells are capable of directly responding to lipopolysaccharide (LPS) and peptidoglycan from periodontal pathogens.
2. Proliferation of GE cells were stimulated with P. gingivalis LPS as well as Echerichia coli LPS, in a dose dependent manner, accompanied by accumulation of IL-1β, IL-6, and TNFα-mRNA.
3. Expression vectors of CD14, TLR-2, TLR-4, and a luciferase reporter plasmid containing κB-site of the mouse ELAM-1 gene have been constructed. GE cells were cotransfected with these plasmids, cultured with or without of LPS, and then luciferase assay was performed. Overexpression of CD-14 and TLR-4 increased the LPS-response of GE cells through the activation of NF-κB, whereas that of TLR-2 did not. These results suggested that CD-14 and TLR-4 could function as receptors for LPS in gingival epithelial cells.
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