Lipopolysaccharide preparation extracted from Porphyromonas gingivalis lipoprotein-deficient mutant shows a marked decrease in Toll-like receptor 2-mediated signaling
Lipopolysaccharide preparation extracted from Porphyromonas gingivalis lipoprotein-deficient mutant shows a marked decrease in Toll-like receptor 2-mediated signaling
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DOI：
10.1128/iai.73.4.2157-2163.2005
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发表时间：
2005-04-01
影响因子：
3.1
通讯作者：
Ogawa, T
中科院分区：
文献类型：
--
作者：
Asai, Y;Hashimoto, M;Ogawa, T
我们最近证明，一种新的 PG1828 编码脂蛋白 (PG1828LP) 能够从牙龈卟啉单胞菌脂多糖 (LPS) 制剂中分离出来，并且我们发现它通过 Toll 样受体 2 (TLR2) 依赖性途径表现出与大肠杆菌 LPS 相似的强细胞活化作用。为了确定 PG1828LP 对细胞激活的毒力，我们使用 ermF-ermAM 抗生素抗性盒通过等位基因交换诱变产生了牙龈卟啉单胞菌菌株 381 的 PG1828 缺陷突变体。来自 PG1828 缺陷突变体 (Delta PG1828-LPS) 的高度纯化的 LIPS 制剂在银染凝胶中显示出与来自野生型菌株 (WT-LPS) 的 LPS 制剂几乎相同的梯状图案，以及与 WT-LPS 制剂相似的鲎变形细胞裂解物活性。然而，Delta PG1828-LPS 制剂激活 TLR2 表达细胞中 NF-κ B 的能力明显减弱。与WT-LPS制剂相比，人牙龈成纤维细胞对Delta PG1828-LPS制剂的细胞因子产生也有所减少，并且其活性与TLR4对牙龈卟啉单胞菌高纯度脂质A的刺激相当。此外，与野生型菌株相比，将PG1828缺陷突变体腹腔注射到小鼠体内后很少观察到致死毒性，而Delta PG1828-LPS制剂则没有表现出致死毒性。综上所述，这些结果清楚地表明 PG1828LP 在炎症反应中发挥重要作用，并且可能是牙龈卟啉单胞菌的主要毒力因子。
We recently demonstrated that a new PG1828-encoded lipoprotein (PG1828LP) was able to be separated from a Porphyromonas gingivalis lipopolysaccharide (LPS) preparation, and we found that it exhibited strong cell activation, similar to that of Escherichia coli LPS, through a Toll-like receptor 2 (TLR2)-dependent pathway. In order to determine the virulence of PG1828LP toward cell activation, we generated a PG1828-deficient mutant of P. gingivalis strain 381 by allelic exchange mutagenesis using an ermF-ermAM antibiotic resistance cassette. A highly purified preparation of LIPS from a PG1828-deficient mutant (Delta PG1828-LPS) showed nearly the same ladder-like patterns in silver-stained gels as a preparation of LPS from a wild-type strain (WT-LPS), as well as Limulus amoebocyte lysate activities that were similar to those of the WT-LPS preparation. However, the ability of the Delta PG1828-LPS preparation to activate NF-kappa B in TLR2-expressing cells was markedly attenuated. Cytokine production by human gingival fibroblasts was also decreased in response to the Delta PG1828-LPS preparation in comparison with the WT-LPS preparation, and the activity was comparable to the stimulation of highly purified lipid A of P. gingivalis by TLR4. Further, lethal toxicity was rarely observed following intraperitoneal injection of the PG1828-deficient mutant into mice compared to that with the wild-type strain, while the Delta PG1828-LPS preparation showed no lethal toxicity. Taken together, these results clearly indicate that PG1828LP plays an essential role in inflammatory responses and may be a major virulence factor of P. gingivalis.