The Mannose-binding Lectin In Immune Responses Against Aspergillus Fumigatus Keratitis In Human Corneal Epithelial Cells

Objective To investigate the expression of MBL in the cultured human cornea epithelial cells(HCECs), and observe the variation of MBL and inflammatory cytokines after the Aspergillus fumigatus spore antigens'stimulation, then analyse the function of MBL in the Aspergillus fumigatus keratitis.Methods When ninety percent of the cells were attached, the medium was replaced with serum-free medium for 24 hours and then the inactivated Aspergillus fumigatus spores were added to the cultures. Harvest the cells after 30 minutes, 1hour,2 hours,4 hours,6 hours,8 hours respectively and then processed for RNA extraction. The expressions of MBL mRNA in the stimulated HCECs were determined by semiquantitative reverse-transcription polymerase chain reaction(RT-PCR). The concentration of MBL,NF-κB,IL-1β,TNF-a,IL-6 and IL-10 in the stimulated cell supernatant were assessed by enzyme-linked immunosorbent assays(ELISA).Results The expressions of MBL in the normal HCECs were very little. The expressions of MBL mRNA in the stimulated HCECs were increased gradually as the stimulation time extended and peaked at the two hours after stimulation; the concentration of the MBL in the HCECs total protein which were measured by ELISA peaked at two hours after stimulation; the concentration of MBL in the cell supernatant were decreased gradually, and the concentration of NF-κB,IL-1β,TNF-α,IL-6 and IL-10 peaked at two hours,one hou,two hours,four hours and six hours after stimulation respectively.Conclusions 1. The HCECs can produced MBL.2.MBL contributes to the recognition of HCECs to the Aspergillus fumigatus spore antigens.3.The MBL may induce the production of the downstream inflammatory cytokines through activating the NF-κB signaling pathway of human corneal epithelium cells, so MBL may play a role in the early innate immunity in the cornea against fungal infection.