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Background  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 Ca 2+  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).    Methods  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).    Results  We demonstrated that an intracellular Ca 2+  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 Ca 2+  signaling and the secretion of IL-6, CXCL1 and TNF-α.    Conclusions  PLCγ2 plays a critical role for Ca 2+  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.

Phospholipase Cγ2 is critical for Ca2+ flux and cytokine production in anti-fungal innate immunity of human corneal epithelial cells