Kawakami Atsushi, Nakashima Koto, Tamai Mami, Nakamura Hideki, Iwanaga Nozomi, Fujikawa Keita, Aramaki Toshiyuki, Arima Kazuhiko, Iwamoto Naoki, Ichinose Kunihiro, Kamachi Makoto, Ida Hiroaki, Origuchi Tomoki, Eguchi Katsumi
First Department of Internal Medicine, Graduate School of Biomedical Sciences, Nagasaki University, Nagasaki, Japan.
J Rheumatol. 2007 May;34(5):1019-26. Epub 2007 Apr 1.
We investigated expression of toll-like receptor (TLR) in labial salivary glands of patients with Sjögren's syndrome (SS) and functional TLR expression in the cultured salivary gland cell line.
Expression of TLR2, TLR3, TLR4, and myeloid differentiation factor 88 (MyD88) in labial salivary glands was examined by immunohistochemistry. Human salivary gland (HSG) cell-line cells were cultured with TLR ligands [peptidoglycan, poly (I:C) and lipopolysaccharide], and CD54 expression and interleukin 6 (IL-6) production was studied. Phosphorylation of extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK), p38, and Akt was examined by Western blotting. Activation of nuclear factor-kappaB (NF-kappaB) p65 in HSG cells was studied by NF-kappaB p65 nuclear translocation by microscopic immunofluorescence or chemiluminescent electrophoretic mobility shift assay and detection of NF-kappaB p65 phosphorylation.
TLR2, TLR3, TLR4, and MyD88 were more strongly expressed in the labial salivary glands of SS patients (n =12) than in control subjects (n = 4), and were found in salivary-infiltrating mononuclear cells as well as acinar cells and ductal epithelial cells. In cultured HSG cells, a similar expression pattern was observed, and TLR ligands stimulated CD54 expression and IL-6 production. TLR ligands induced phosphorylation of ERK, JNK, and p38 in HSG cells, but not Akt phosphorylation or activation of NF-kappaB p65.
Although the putative ligands remain to be determined, our study indicated the activation of the TLR-mediated immune response in SS, and suggested that the TLR effect is mediated through the mitogen-activated protein kinase pathway.
我们研究了干燥综合征（SS）患者唇腺中Toll样受体（TLR）的表达以及培养的涎腺细胞系中功能性TLR的表达。
采用免疫组织化学法检测唇腺中TLR2、TLR3、TLR4和髓样分化因子88（MyD88）的表达。用人涎腺（HSG）细胞系细胞与TLR配体[肽聚糖、聚（I:C）和脂多糖]进行培养，研究CD54表达和白细胞介素6（IL-6）的产生。通过蛋白质印迹法检测细胞外信号调节激酶（ERK）、c-Jun氨基末端激酶（JNK）、p38和Akt的磷酸化。通过显微免疫荧光或化学发光电泳迁移率变动分析以及检测NF-κB p65磷酸化来研究HSG细胞中核因子-κB（NF-κB）p65的激活。
TLR2、TLR3、TLR4和MyD88在SS患者（n = 12）的唇腺中表达比对照组（n = 4）更强，且在涎腺浸润的单核细胞以及腺泡细胞和导管上皮细胞中均有发现。在培养的HSG细胞中观察到类似的表达模式，并且TLR配体刺激了CD54表达和IL-6的产生。TLR配体诱导了HSG细胞中ERK、JNK和p38的磷酸化，但未诱导Akt磷酸化或NF-κB p65的激活。
尽管假定的配体仍有待确定，但我们的研究表明SS中TLR介导的免疫反应被激活，并提示TLR效应是通过丝裂原活化蛋白激酶途径介导的。
