Induction of Porphyromonas gingivalis GroEL signaling via binding to Toll-like receptors 2 and 4
Induction of Porphyromonas gingivalis GroEL signaling via binding to Toll-like receptors 2 and 4
复制标题
DOI：
10.1111/j.1399-302x.2006.00286.x
复制
发表时间：
2006-08-01
影响因子：
--
通讯作者：
Hanazawa, S.
中科院分区：
文献类型：
--
作者：
Argueta, J. G. M.;Shiota, S.;Hanazawa, S.
背景/目的：热休克蛋白 60 (HSP60) 已被认为是感染性和自身免疫性疾病中的重要分子。尽管牙龈卟啉单胞菌 GroEL（HSP60 的同源物）是炎症细胞因子的有效刺激剂，但其受体和信号传导机制尚未详细了解。在本研究中，我们研究了 Toll 样受体 (TLR) 家族是否作为牙龈卟啉单胞菌 GroEL 受体发挥功能性作用。方法：使用人巨噬细胞样 THP-1 细胞，并通过荧光素酶测定测量重组牙龈卟啉单胞菌 GroEL 刺激的细胞的核因子 -κ B (NF-κ B) 活性。使用流式细胞术分析确定异硫氰酸荧光素 (FITC) 标记的 GroEL 与 THP-1 细胞的结合。此外，使用抗人TLR（抗hTLR）2和抗hTLR4单克隆抗体来评估TLR2和TLR4作为GroEL受体的功能作用。结果：我们通过荧光素酶测定观察到纯化的重组GroEL能够刺激THP-1细胞中的NF-kappa B转录活性。流式细胞术分析表明，FITC 标记的 GroEL 以剂量依赖性方式与 THP-1 细胞结合。我们对 FITC 标记和未标记的 GroEL 进行的结合竞争分析表明，它以特定的作用模式与细胞结合。另一方面，GroEL 刺激的 NF-κ B 转录活性被抗 hTLR2 和抗 hTLR4 抗体显着抑制，并且两种抗体组合的抑制更强烈。结论：我们目前的研究表明，牙龈卟啉单胞菌 GroEL 通过 TLR2 或 TLR4 以及通过组合在 THP-1 细胞中诱导其细胞内信号级联反应。 两个受体的。
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-kappa B (NF-kappa B) 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-kappa B 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-kappa B 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.