論文 - 詳細
| RRC ID | 42702 |
|---|---|
| 著者 | Argueta JG, Shiota S, Yamaguchi N, Masuhiro Y, Hanazawa S. |
| タイトル | Induction of Porphyromonas gingivalis GroEL signaling via binding to Toll-like receptors 2 and 4. |
| ジャーナル | Oral Microbiol Immunol |
| Abstract |
BACKGROUND/AIMS:Heat shock protein 60 (HSP60) has been recognized as an important molecule in infectious and autoimmune diseases. Although Porphyromonas gingivalis GroEL, a homologue of HSP60, is a potent stimulator of inflammatory cytokines, its receptor and signaling mechanisms are not yet understood in detail. In this study, we investigated whether the Toll-like receptor (TLR) family plays a functional role as a P. gingivalis GroEL receptor. METHODS:Human macrophage-like THP-1 cells were used and the nuclear factor-kappaB (NF-kappaB) activity of cells stimulated with a recombinant P. gingivalis GroEL was measured with a luciferase assay. Flow cytometry analysis was used to determine the binding to THP-1 cells of fluorescein isothiocyanate (FITC)-labeled GroEL. In addition, anti-human TLR (anti-hTLR)2 and anti-hTLR4 monoclonal antibodies were used to assess the functional role of TLR2 and TLR4 as the receptors for GroEL. RESULTS:We observed by luciferase assay that the purified recombinant GroEL was able to stimulate NF-kappaB transcriptional activity in THP-1 cells. Flow cytometry analysis showed that the FITC-labeled GroEL bound to THP-1 cells in a dose-dependent fashion. Our binding competition analysis with FITC-labeled and unlabeled GroEL showed that it bound to the cells as a specific mode of action. On the other hand, GroEL-stimulated NF-kappaB transcriptional activity was significantly inhibited by anti-hTLR2 and anti-hTLR4 antibodies and was inhibited more strongly by a combination of both antibodies. CONCLUSION:Our present study demonstrates that P. gingivalis GroEL induces its intracellular signaling cascade in THP-1 cells via TLR2 or TLR4 and via a combination of both receptors. |
| 巻・号 | 21(4) |
| ページ | 245-51 |
| 公開日 | 2006-8-1 |
| DOI | 10.1111/j.1399-302X.2006.00286.x |
| PII | OMI286 |
| PMID | 16842509 |
| MeSH | Antibodies, Monoclonal Chaperonin 60 / pharmacology Chaperonin 60 / physiology* Flow Cytometry Fluorescein-5-isothiocyanate Fluorescent Dyes Humans Lipopolysaccharides / pharmacology Luciferases Luminescent Agents Macrophages / microbiology Monocytes / microbiology NF-kappa B / physiology Porphyromonas gingivalis / physiology* Protein Binding / drug effects Protein Binding / physiology* Signal Transduction / drug effects Signal Transduction / physiology* Toll-Like Receptor 2 / drug effects Toll-Like Receptor 2 / physiology* Toll-Like Receptor 4 / drug effects Toll-Like Receptor 4 / physiology* Transcription, Genetic / genetics |
| 引用数 | 24 |
| WOS 分野 | DENTISTRY, ORAL SURGERY & MEDICINE IMMUNOLOGY MICROBIOLOGY |
| リソース情報 | |
| ヒト・動物細胞 | THP-1 |