Yamaguchi Ryusuke, Yoshimura Atsutoshi, Yoshioka Hidenobu, Kaneko Takashi, Hara Yoshitaka
Department of Periodontology, Nagasaki University Graduate School of Biomedical Sciences, Nagasaki, Japan.
J Periodontol. 2009 Mar;80(3):512-20. doi: 10.1902/jop.2009.080393.
In our previous study, we found that the ability of supragingival plaque to induce Toll-like receptor (TLR)4-mediated stimulation was positively associated with plaque score and bleeding on probing (BOP) at the sampled sites and that the ability to induce TLR2-mediated stimulation was negatively associated with probing depth (PD) and clinical attachment level (CAL). Because signaling from TLR leads to the induction of pro- and anti-inflammatory cytokines, we further analyzed the influence of the ability of supragingival plaque to induce TLR2-/TLR4-mediated stimulation of cytokine production by peripheral blood mononuclear cells (PBMCs).
The abilities of 125 plaque samples to induce TLR2- or TLR4-mediated stimulation were determined using genetically engineered Chinese hamster ovary reporter cells that express a reporter molecule upon activation of nuclear factor-kappa B through TLR2 or TLR4. PBMCs were stimulated with each plaque sample, and the production of proinflammatory cytokines (tumor necrosis factor-alpha and interleukin [IL]-6 and -8) and an anti-inflammatory cytokine (IL-10) was analyzed by enzyme-linked immunosorbent assay.
The levels of the cytokines produced by PBMCs all correlated with the ability of supragingival plaque to induce TLR4-mediated stimulation but not with its ability to induce TLR2-mediated stimulation. Cytokine production was inhibited by an anti-TLR4 monoclonal antibody and a TLR4 antagonist, compound 406. The levels of cytokines were associated with plaque index, BOP, PD, and CAL at the sampled sites.
The production of pro-/anti-inflammatory cytokines by PBMCs was associated with the ability of supragingival plaque to induce TLR4-mediated stimulation. The cytokines induced by supragingival plaque via TLR4 might modulate periodontal status.
在我们之前的研究中，我们发现龈上菌斑诱导Toll样受体（TLR）4介导刺激的能力与采样部位的菌斑评分和探诊出血（BOP）呈正相关，而诱导TLR2介导刺激的能力与探诊深度（PD）和临床附着水平（CAL）呈负相关。由于TLR信号传导会导致促炎和抗炎细胞因子的诱导，我们进一步分析了龈上菌斑诱导外周血单核细胞（PBMC）产生TLR2/TLR4介导的细胞因子刺激能力的影响。
使用经基因工程改造的中国仓鼠卵巢报告细胞来测定125个菌斑样本诱导TLR2或TLR4介导刺激的能力，这些细胞在通过TLR2或TLR4激活核因子-κB时会表达一种报告分子。用每个菌斑样本刺激PBMC，并通过酶联免疫吸附测定法分析促炎细胞因子（肿瘤坏死因子-α和白细胞介素[IL]-6和-8）和抗炎细胞因子（IL-10）的产生。
PBMC产生的细胞因子水平均与龈上菌斑诱导TLR4介导刺激的能力相关，而与其诱导TLR2介导刺激的能力无关。细胞因子的产生受到抗TLR4单克隆抗体和TLR4拮抗剂化合物406的抑制。细胞因子水平与采样部位的菌斑指数、BOP、PD和CAL相关。
PBMC产生促炎/抗炎细胞因子与龈上菌斑诱导TLR4介导刺激的能力相关。龈上菌斑通过TLR4诱导的细胞因子可能会调节牙周状态。
