Toll-like receptor expression in human keratinocytes:: Nuclear factor κB controlled gene activation by Staphylococcus aureus is toll-like receptor 2 but not toll-like receptor 4 or platelet activating factor receptor dependent
Toll-like receptor expression in human keratinocytes:: Nuclear factor κB controlled gene activation by Staphylococcus aureus is toll-like receptor 2 but not toll-like receptor 4 or platelet activating factor receptor dependent
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DOI：
10.1111/j.1523-1747.2003.12630.x
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发表时间：
2003-12-01
影响因子：
6.5
通讯作者：
Ollert, M
中科院分区：
文献类型：
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作者：
Mempel, M;Voelcker, V;Ollert, M
对原代培养的人角质形成细胞进行Toll样受体(TLR)家族各成员的表达筛选。聚合酶链式反应显示角质形成细胞表达TLR1、TLR2、TLR3、TLR5和TLR9，但不表达TLR4、TLR6、TLR7、TLR8和TLR10。免疫组织化学也证实了葡萄球菌化合物信号转导的关键受体TLR2的表达，而TLR4则呈阴性染色。接下来，我们分析了金黄色葡萄球菌8325-4株对促炎症核转录因子kappaB的激活作用。利用核抽提物凝胶移位、RELA染色和荧光素酶报告基因载体，我们发现该细菌明显诱导了核因子kappaB的易位。这种易位诱导了核因子kappaB控制的基因的转录，如诱导型一氧化氮合酶、COX2和白细胞介素8。在这些基因转录之后，白介素8蛋白和NO的产量增加。用单抗和特异性的血小板活化因子受体抑制剂CV3988进行的抑制实验表明，金黄色葡萄球菌对核因子kappaB的激活依赖于TLR2，而不依赖于TLR4或血小板激活因子受体。同时，纯化的葡萄球菌细胞壁成分脂磷壁酸和肽聚糖，已知通过TLR2发出信号，也在人角质形成细胞中显示核因子kappaB易位，表明葡萄球菌细胞壁在人角质形成细胞的天然免疫刺激中起着至关重要的作用。这些结果有助于解释金黄色葡萄球菌及其细胞壁成分在各种皮肤炎症性疾病中对人角质形成细胞的复杂激活。
Cultured primary human keratinocytes were screened for their expression of various members of the toll-like receptor (TLR) family. Keratinocytes were found to constitutively express TLR1, TLR2, TLR3, TLR5, and TLR9 but not TLR4, TLR6, TLR7, TLR8, or TLR10 as shown by polymerase chain reaction analysis. The expression of the crucial receptor for signaling of staphylococcal compounds TLR2 was also confirmed by immunohistochemistry, in contrast to TLR4, which showed a negative staining pattern. Next, we analyzed the activation of the proinflammatory nuclear transcription factor kappaB by Staphylococcus aureus strain 8325-4. Using nuclear extract gel shifts, RelA staining, and luciferase reporter transfection plasmids we found a clear induction of nuclear factor kappaB translocation by the bacteria. This translocation induced the transcription of nuclear factor kappaB controlled genes such as inducible nitric oxide synthetase, COX2, and interleukin-8. Transcription of these genes was followed by production of increased amounts of interleukin-8 protein and NO. Inhibition experiments using monoclonal antibodies and the specific platelet activating factor receptor inhibitor CV3988 showed that nuclear factor kappaB activation by S. aureus was TLR2 but not TLR4 or platelet activating factor receptor dependent. In line, the purified staphylococcal cell wall components lipoteichoic acid and peptidoglycan, known to signal through TLR2, also showed nuclear factor kappaB translocation in human keratinocytes, indicating a crucial role of the staphylococcal cell wall in the innate immune stimulation of human keratinocytes. These results help to explain the complex activation of human keratinocytes by S. aureus and its cell wall components in various inflammatory disorders of the skin.