Fungus induces the release of IL-8 in human corneal epithelial cells, via Dectin-1-mediated protein kinase C pathways
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Supported by National Natural Science Foundation of China (No. 81470609; No. 81170825); the Specialized Research Fund for the Doctoral Program of Higher Education (No. 20123706110003); the Key Project of Natural Science Foundation of Shandong Province (No. ZR2012FZ001); the Youth Project of Natural Science Foundation of Shandong Province (No. ZR2013HQ007).

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    Abstract:

    AIM: To identify whether Aspergillus fumigatus (A. fumigatus) hyphae antigens induced the release of interleukin-8 (IL-8) in anti-fungal innate immunity of cultured human corneal epithelial cells (HCECs) and determine the involvement of intracellular signalling pathways.METHODS: HCECs were treated with A. fumigatus hyphae antigens with different concentrations and time. The cytoplasmic calcium of HCECs were assessed by fluorescence imaging. Western blot was used to detect the expression of Ca2+-dependent protein kinase C (PKC). The IL-8 levels were determined by specific human IL-8 enzyme-linked immunosorbent assay (ELISA) and reverse transcriptase polymerase chain reaction (RT-PCR). Using a series of pharmacological inhibitors, we examined the upstream signalling pathway responsible for IL-8 expression in response to A. fumigatus hyphae antigens.RESULTS: Cells exposed to A. fumigatus hyphae antigens showed higher level of IL-8 mRNA expression and protein production. We demonstrated here that stimulation of HCECs with A. fumigatus hyphae triggers an intracellular Ca2+ flux and results in the activation of Ca2+-dependent PKC (α, βⅠand βⅡ) which can be attenuated by pre-treatment of cells with laminarin, suggesting that Dectin-1 signals pathway induced cytoplasmic calcium and influence the activation of PKC in HCECs. Inhibitors of Ca2+-dependent PKC (Ro-31-8220 and Go6976) significantly abolished hyphae-induced expression of IL-8.CONCLUSION: Our findings suggest that A. fumigatus hyphae-induced IL-8 expression was regulated by the activation of Dectin-1-mediated Ca2+-dependent PKC in HCECs.

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Xu-Dong Peng, Gui-Qiu Zhao, Jing Lin, et al. Fungus induces the release of IL-8 in human corneal epithelial cells, via Dectin-1-mediated protein kinase C pathways. Int J Ophthalmol, 2015,8(3):441-447

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Publication History
  • Received:November 03,2014
  • Revised:January 05,2015
  • Adopted:January 05,2015
  • Online: June 09,2015
  • Published: