Pneumolysin-induced CXCL8 production by nasopharyngeal epithelial cells is dependent on calcium flux and MAPK activation via Toll-like receptor 4
Pneumolysin-induced CXCL8 production by nasopharyngeal epithelial cells is dependent on calcium flux and MAPK activation via Toll-like receptor 4
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DOI：
10.1016/j.micinf.2010.10.003
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发表时间：
2011-01-01
影响因子：
5.8
通讯作者：
Murdoch, Craig
中科院分区：
文献类型：
--
作者：
Dogan, Semih;Zhang, Qibo;Murdoch, Craig
肺炎链球菌的自然生态位是鼻咽粘膜，并且肺炎球菌的鼻咽携带广泛流行。溶血素（Ply）是由链球菌产生的一种成孔蛋白。肺炎，可能是重要的驱动先天免疫反应的鼻咽。我们研究了Ply诱导鼻咽癌细胞产生CXCL 8，并进一步分析了这种诱导的机制。用来源于Ply缺陷型、野生型肺炎球菌和重组Ply的细菌培养物的上清液刺激底特律鼻咽细胞，并通过ELISA测量CXCL 8。使用特异性抑制剂分析MAP激酶家族成员在Ply诱导的CXCL 8产生中的作用，通过免疫印迹测量NF-κ B活性，并通过CXCL 8启动子荧光素酶测定分析Ply介导的TLR 4活化。Ply显著增加底特律和原代鼻细胞中CXCL 8的产生。流式细胞仪分析显示，底特律细胞表达细胞表面TLR 4。CXCL 8的产生依赖于细胞内Ca 2+水平的变化，也依赖于NF-κ B B通过激活TLR 4和MAP激酶信号传导的变化。Ply通过涉及Ca 2+动员和MAPK和NF-κ B通过TLR 4活化的信号传导机制诱导鼻咽细胞产生CXCL 8。这可能是重要的，在调节鼻咽免疫肺炎球菌定植。(C)2010年巴斯德研究所。由Elsevier Masson SAS出版。All rights reserved.
The natural niche of Streptococcus pneumoniae is the nasopharyngeal mucosa and nasopharyngeal carriage of pneumococci is widely prevalent. Pneumolysin (Ply), a pore-forming protein produced by S. pneumonia, may be important in driving the innate immune response of the nasopharynx. We studied the Ply-induced production of CXCL8 by nasopharyngeal cells and further analysed the mechanism of this induction. Detroit nasopharyngeal cells were stimulated with supernatants derived from bacterial cultures of Ply-deficient, wild-type pneumococci and recombinant Ply, and CXCL8 measured by ELISA. The role of MAP kinase family members in Ply-induced CXCL8 production was analysed using specific inhibitors, NF-kappa B activity was measured by immunoblot and Ply-mediated TLR4 activation analysed by a CXCL8 promotor luciferase assay. Ply significantly increased production of CXCL8 in Detroit and primary nasal cells. Flow cytometric analysis showed that Detroit cells express cell surface TLR4. CXCL8 production was dependent on changes in the intracellular Ca2+ levels and also by NF-kappa B via activation of TLR4, and MAP kinase signalling. Ply induces production of CXCL8 by nasopharyngeal cells using signalling mechanisms involving Ca2+ mobilisation and activation of MAPK and NF-kappa B via TLR4. This may be important in regulating nasopharyngeal immunity against pneumococcal colonization. (C) 2010 Institut Pasteur. Published by Elsevier Masson SAS. All rights reserved.