Protease-activated receptor 2 mediates human beta-defensin 2 and CC chemokine ligand 20 mRNA expression in response to proteases secreted by Porphyromonas gingivalis
Protease-activated receptor 2 mediates human beta-defensin 2 and CC chemokine ligand 20 mRNA expression in response to proteases secreted by Porphyromonas gingivalis
复制标题
DOI：
10.1128/iai.00455-07
复制
发表时间：
2007-09-01
影响因子：
3.1
通讯作者：
Dale, Beverly A.
中科院分区：
文献类型：
--
作者：
Dommisch, Henrik;Chung, Whasun O.;Dale, Beverly A.
口腔病原体牙龈卟啉单胞菌分泌蛋白酶，例如精氨酸牙龈蛋白酶 B (RgpB)，可激活蛋白酶激活受体 (PAR)。人β-防御素 (hBD) 和上皮细胞产生的巨噬细胞炎症蛋白 3α/CC 趋化因子配体 20 (CCL20) 是抗菌肽，提供细胞因子功能并在先天免疫中发挥重要作用。本研究的目的是确定当暴露于牙龈卟啉单胞菌无细胞培养物上清液或纯化的 RgpB 时，PAR 家族的特定成员是否介导牙龈上皮细胞 (GEC) 中这些先天免疫标志物的表达。 GEC 中的 hBD-2 mRNA 是响应上清液和来自牙龈卟啉单胞菌的纯化 RgpB 进行诱导的（分别为 P = 0.02 和 P = 0.016）。这种效应被蛋白酶抑制剂甲苯磺酰-L-赖氨酸氯甲基酮 (TLCK) 消除（P < 0.05）。响应牙龈卟啉单胞菌上清液和纯化的RgpB，在PAR-2基因敲低细胞中hBD-2 mRNA表达显着降低，而在PAR-1基因敲低细胞中未检测到变化。 CCL20 mRNA 表达也因牙龈卟啉单胞菌上清液而增加，并且这种效应被蛋白酶抑制剂 TLCK 阻断（分别为 P = 0.05 和 P = 0.024），并且在 PAR-2 基因敲除细胞中也被阻断。我们的数据表明，牙龈卟啉单胞菌上清液和纯化的 RgpB 上调的 hBD-2 和 CCL20 mRNA 是通过 PAR-2 介导的，而不是通过 PAR-1 介导的，并且蛋白酶在 GEC 中先天免疫反应的调节中发挥作用。 GEC 使用 PAR 来识别牙龈卟啉单胞菌并介导参与先天免疫的细胞反应。
The oral pathogen Porphyromonas gingivalis secretes proteases such as Arg-gingipain B (RgpB) that activate protease-activated receptors (PARs). Human beta-defensins (hBDs) and the macrophage inflammatory protein 3 alpha/CC chemokine ligand 20 (CCL20) produced by epithelial cells are antimicrobial peptides that provide cytokine function and play an important role in innate immunity. The aim of the present study was to determine whether specific members of the PAR family mediate the expression of these innate immunity markers in gingival epithelial cells (GECs) when exposed to P. gingivalis cell-free culture supernatant or purified RgpB. hBD-2 mRNA in GECs was induced in response to supernatant and purified RgpB from P. gingivalis (P = 0.02 and P = 0.016, respectively). This effect was abrogated by the protease inhibitor tosyl-L-lysine chloromethyl ketone (TLCK) (P < 0.05). In response to P. gingivalis supernatant and to purified RgpB, the hBD-2 mRNA expression was significantly decreased in PAR-2 gene knockdown cells, whereas no change was detected in PAR-I gene knockdown cells. CCL20 mRNA expression also increased in response to the supernatant of P. gingivalis, and this effect was blocked by the protease inhibitor, TLCK (P = 0.05 and P = 0.024, respectively), and was blocked in PAR-2 gene knockdown cells. Our data indicate that hBD-2 and CCL20 mRNA up-regulation by P. gingivalis supernatant and purified RgpB was mediated via PAR-2, but not via PAR-1, and that proteases play a role in the regulation of innate immune responses in GECs. GECs use PARs to recognize P. gingivalis and mediate cell responses involved in innate immunity.