Various members of the Toll-like receptor family contribute to the innate immune response of human epidermal keratinocytes

被引:288
作者
Köllisch, G
Kalali, BN
Voelcker, V
Wallich, R
Behrendt, H
Ring, J
Bauer, S
Jakob, T
Mempel, M
Ollert, M
机构
[1] Tech Univ Munich, Dept Dermatol & Allergy, D-80802 Munich, Germany
[2] Tech Univ Munich, Dept Med Microbiol Immunol & Hyg, D-80802 Munich, Germany
[3] Tech Univ Munich, Clin Res Div Mol & Clin Allergotox, D-80802 Munich, Germany
[4] Heidelberg Univ, Inst Immunol, D-6900 Heidelberg, Germany
[5] GSF, Natl Res Ctr Environm & Hlth, Div Environm Dermatol & Allergy GSF TUM, Neuherberg, Germany
关键词
innate immunity of the skin; HaCaT cells; LPS; NF-kappa B;
D O I
10.1111/j.1365-2567.2005.02122.x
中图分类号
R392 [医学免疫学]; Q939.91 [免疫学];
学科分类号
100102 ;
摘要
Toll-like receptors (TLRs) are important pattern recognition molecules that activate the nuclear factor (NF)-kappa B pathway leading to the production of antimicrobial immune mediators. As keratinocytes represent the first barrier against exogenous pathogens in human skin, we investigated their complete functional TLR1-10 expression profile. First, reverse transcription-polymerase chain reaction (PCR) analysis revealed a very similar pattern of TLR mRNA expression when comparing freshly isolated human epidermis and cultured primary human keratinocytes. Thus, further experiments were carried out with primary keratinocytes in comparison with the spontaneously immortalized human keratinocyte cell line HaCaT. The quantitative expression of TLR1-10 mRNA in real-time PCR of primary human keratinocytes and HaCaT cells was analysed. Both cell types constitutively expressed TLR2, TLR3, TLR5, and to a lesser extent TLR10. TLR4 was only found in HaCaT cells, TLR1 to a higher degree in primary keratinocytes. In line with this, LPS induced mRNA expression of CD14 and TLR4 only in HaCaT cells. After stimulation with various TLR ligands, the NF-kappa B-activated chemokine interleukin-8 (IL-8) was measured. In primary keratinocytes and HaCaT cells the TLR3 ligand poly (I:C) was the most potent stimulator of IL-8 secretion. The TLR ligands peptidoglycan, Pam(3)Cys and flagellin which bind to TLR2, TLR1/TLR2 heterodimer, and TLR5, respectively, also induced IL-8 secretion, whereas no IL-8 was induced by LPS, R-848, loxoribine and cytosine guanine dinucleotide-containing oligodeoxynucleotide. A corresponding pattern was found in the RelA NF-kappa B translocation assay after ligand stimulation of primary keratinocytes. These studies provide substantial evidence for a functional TLR expression and signalling profile of normal human keratinocytes contributing to the antimicrobial defence barrier of human skin.
引用
收藏
页码:531 / 541
页数:11
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