Activition Of Dectin-1 In Aspergillus Fumigatus-Induced Innate Immune Responses In Human Cornea Epithelial Cells

Objective To investigate expression and role of Dectin-1 and its corresponding factors in human corneal epithelial cells exposed to Aspergillus fumigatus antigens.Methods Stimulated cultured human corneal epithelium cells with Aspergillus fumigatus antigens were collected and expression of Dectin-1 mRNA was done by RT-PCR; expression of Dectin-1 in protein level was done by immunocytochemistry SP method and ELISA; level of expression of relative downstream factors like NF-κB, TNF-a,IL-1β, IL-10 and CXCL1 in cell culture media were measured by ELISA. The comparisons among groups were done by SPSS 17.0 software.Results Dectin-1 mRNA existed in normal HCECs and increased after fungal stimulation,and its expression upregulated by stimulation time,reaching their peak at 2h,and every stimulated groups were statistically significant compared with blank group(P=0.000). Expression of Dectin-1 in protein level were successfully detected by immunocytochemistry SP method in every groups,and results by ELISA were a litte different from mRNA,for at 0.5h Dectin-1 decreased while at 1h increased again, peaked at 2h and then dropped,and every stimulated groups were statistically significant compared with blank group(P=0.000). NF-κB in cell culture media increased after fungal stimulation and peaked at 2h,and so did TNF-a(peaked at 2h),IL-1β(peaked at lh)and CXCL1 (peaked at 4h),while IL-10 had different change(peaked at 6h) for its reverse role in fungal infection.And for all of them every stimulated groups were statistically significant compared with blank group(P=0.000).Conclusion Our findings indicate that human corneal epithelial cells has functional Dectin-1 and activation of Dectin-1 may play a critical role in pathogenesis of keratomycosis.