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- Title
Phospholipase Cγ2 is critical for Ca<sup>2+</sup> flux and cytokine production in anti-fungal innate immunity of human corneal epithelial cells.
- Authors
Peng, Xudong; Zhao, Guiqiu; Lin, Jing; Qu, Jianqiu; Zhang, Yingxue; Li, Cui
- Abstract
<bold>Background: </bold>Fungal keratitis (FK) is a sight-threatening disease, accounting for a significant portion with its complex presentation, suboptimal efficacy of the existing therapies and uncontrollable excessive innate inflammation. Phospholipase C-γ2 (PLCγ2) is a non-receptor tyrosine kinase that plays an important role at the early period of innate immunity. This study aimed to identify the role of PLCγ2 in Dectin-1-mediated Ca2+ Flux and its effect on the expression of proinflammatory mediators at the exposure to Aspergillus fumigatus (A. fumigatus) hyphae antigens in human corneal epithelial cells (HCECs).<bold>Methods: </bold>The HCECs were preincubated with or without different inhibitors respectively before A. fumigatus hyphae stimulation. Intracellular calcium flux in HCECs and levels of PLCγ2 and spleen-tyrosine kinase (Syk) were detected by fluorescence imaging and Western Blotting. The expression of proinflammatory mediators was determined by reverse transcriptase polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay (ELISA).<bold>Results: </bold>We demonstrated that an intracellular Ca2+ flux in HCECs was triggered by A. fumigatus hyphae and could be reduced by pre-treatment with PLCγ2-inhibitor U73122. A. fumigatus hyphae induced PLCγ2 phosphorylation was regulated by Dectin-1 via Syk. Furthermore, PLCγ2-deficient HCECs showed a drastic impairment in the Ca2+ signaling and the secretion of IL-6, CXCL1 and TNF-α.<bold>Conclusions: </bold>PLCγ2 plays a critical role for Ca2+ Flux in HCECs stimulated by A. fumigatus hyphae. Syk acts upstream of PLCγ2 in the Dectin-1 signaling pathway. The expressions of proinflammatory mediators induced by A. fumigatus are regulated by the activation of Dectin-1-mediated PLCγ2 signaling pathway in HCECs.
- Subjects
PHOSPHOLIPASES; FUNGAL keratitis; NATURAL immunity; CORNEA; EPITHELIAL cells; PROTEIN-tyrosine kinases; CYTOKINES; CALCIUM metabolism; ASPERGILLUS; CELL culture; CELLULAR signal transduction; ENZYME-linked immunosorbent assay; EPITHELIUM; EYE infections; GENES; IMMUNITY; MYCOSES; POLYMERASE chain reaction; RESEARCH funding; RNA; WESTERN immunoblotting
- Publication
BMC Ophthalmology, 2018, Vol 18, Issue 1, pN.PAG
- ISSN
1471-2415
- Publication type
journal article
- DOI
10.1186/s12886-018-0847-6