The role of RNase 7 in innate cutaneous defense against Pseudomonas aeruginosa
The role of RNase 7 in innate cutaneous defense against Pseudomonas aeruginosa
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DOI：
10.1111/exd.13166
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发表时间：
2017-03-01
影响因子：
3.6
通讯作者：
Harder, Juergen
中科院分区：
文献类型：
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作者：
Rademacher, Franziska;Simanski, Maren;Harder, Juergen
核糖核酸酶RNase7是一种主要来源于皮肤的人类抗菌蛋白，在角质形成细胞中表达。在这里，我们展示了革兰氏阴性杆菌铜绿假单胞菌分泌因子(S)，诱导了人原代角质形成细胞的RNase7基因和蛋白的表达。金属蛋白酶抑制剂marimastat、表皮生长因子受体(EGFR)抑制剂AG1478和EGFR阻断抗体西妥昔单抗显著减弱这种诱导，表明EGFR在铜绿假单胞菌介导的RNase7诱导中起重要作用。与此相一致的是，siRNA介导的ADAM17的下调，减少了铜绿假单胞菌介导的角质形成细胞中RNase7的诱导。ADAM17是一种已知的蛋白水解性金属蛋白酶，介导了可溶性EGFR配体的释放。EGFR的影响也在人体3D皮肤等效物上得到了证明，阻断EGFR可以减少铜绿假单胞菌对RNase7的诱导。阻断Toll样受体5(TLR5)，一种已知由铜绿假单胞菌激活的模式识别受体(PRR)，仅适度降低铜绿假单胞菌介导的角质形成细胞RNase7的诱导。通过中和RNase7的抗菌活性的抗体，证明了RNase7参与皮肤防御铜绿假单胞菌的功能。这些抗体显著抑制了人角质层皮肤提取物控制铜绿假单胞菌生长的能力。综上所述，我们的结果表明，铜绿假单胞菌以EGFR依赖的方式诱导角质形成细胞中RNase7的表达。促进核糖核酸酶7的释放有助于控制铜绿假单胞菌的皮肤生长。
The ribonuclease RNase 7 is a major skin-derived human antimicrobial protein expressed in keratinocytes. Here we show that the gram-negative pathogen Pseudomonas aeruginosa secretes factor(s) that induced RNase 7 gene and protein expression in human primary keratinocytes. The metalloprotease inhibitor marimastat, the epidermal growth factor receptor (EGFR) inhibitor AG1478 and the EGFR blocking antibody cetuximab significantly attenuated this induction, indicating an important role of the EGFR for the P. aeruginosa-mediated RNase 7 induction. In line with this, siRNA-mediated downregulation of ADAM17, a metalloprotease known to proteolytically mediate the release of soluble EGFR ligands, decreased the P. aeruginosa-mediated RNase 7 induction in keratinocytes. The impact of the EGFR was also demonstrated in a human 3D skin equivalent where blockade of the EGFR diminished induction of RNase 7 by P. aeruginosa. Blockade of Toll-like receptor 5 (TLR5), a pattern recognition receptor (PRR) known to be activated by P. aeruginosa, only moderately reduced the P. aeruginosa-mediated RNase 7 induction in keratinocytes. The functional relevance of RNase 7 to participate in cutaneous defense against P. aeruginosa was demonstrated by antibodies that neutralized the antimicrobial activity of RNase 7. These antibodies significantly inhibited the capacity of human stratum corneum skin extracts to control growth of P. aeruginosa. Taken together, our results indicate that P. aeruginosa induces the expression of RNase 7 in keratinocytes in an EGFR-dependent manner. Enhanced release of RNase 7 contributes to control cutaneous growth of P. aeruginosa.