Activition Of Surfactant Protein A And D In Aspergillus Fumigatus-Induced Innate Immune Responses In Human Cornea Epithelial Cells

Objective:To investigate expression and roles of surfactant protein A and D (SP-A,SP-D) and its corresponding factors in tolerated human corneal epithelial cells exposed to aspergillus fumigatus antigens.Methods: Tolerated human corneal epithelium cells(THCEs) were cultured in vitro. Stimulated different groups with aspergillus fumigatus antigens which we maked ourselves at 0.5 hour,1 hour,2 hours,4 hours,6 hours and 8 hours (n=6).Expressions of SP-A and SP-D mRNA was evaluated by Reversed-transcript PCR(RT-PCR); expressions of SP-A and SP-D in proteins'levels was shown by ELISA and immunohistochemistry SP methods; level of expression of NF-κB and relative downstream factors such as TNF-a,IL-1β,IL-8 and IL-10 in supernatant fluid were measured by ELISA also.Results:Both two kinds of mRNA and proteins of SP-A,SP-D were detected in untreated groups.Expressions of SP-A,SP-D and the relative downstream factors raised after being stimulated with aspergillus fumigatus antigens. Expressions of SP-A and SP-D mRNA began to rise at 0.5h groups,which 1.29 and 1.02 times higher than untreated groups (P<0.01); SP-A mRNA and SP-D mRNA were detected the most significantly higher in 2 hours groups (P<0.01).The proteins of SP-A,SP-D began to rise at 30 min groups. There were significances in statistics(P<0.01).NF-kB was activated after treated by aspergillus fumigatus antigens and its changes seemed to have some correlation with SP-A and SP-D.After given fumigatus antigens 1 hour later, TNF-a,IL-1β,IL-8 and IL-10 began to rise and were 1.82,1.43,1.12 and 1.28 times higher than the untreated groups separately(P<0.05). TNF-a and IL-1βat 2 hours groups were the highest, separately 2.80,2.86 times than the untreated groups(P<0.01).There were SP-A and SP-D in untreated THCEs groups. SP-A and SP-D were mainly located in the cytoplasma.And they could crease after being treared with the aspergillus fumigatus antigens.Conclusion:Our findings indicated that human corneal epithelial cells exist SP-A and SP-D and they may play critical roles in pathogenesis of keratomycosis.