| Summary: | 碩士 === 中山醫學大學 === 毒理學研究所 === 91 === In this study, we expressed and purified the fungal immunomodulatory protein (FIP-gts) in E.coli. Using PI staining and flow cytometry, we found that FIP-gts promoted cell cycle progression from G0/G1 phase to S phase in HPBMCs. Pre-treatment of LY294002, a specific inhibitor of PI 3-kinase, was abolished the increased S phase induced by FIP-gts. Analyzing the cytokines induced by FIP-gts, IFN-γ persistent expressed significantly. Pre-treatment of LY294002 was also inhibited the secretion of IFN-γ stimulated by FIP-gts in HPBMCs. Moreover our results demonstrated that Akt, a downstream effecter of phosphatidylinositol 3-kinase (PI 3-kinase), was phosphorylated and activated by FIP-gts. It indicated that PI 3-kinase play an important role in the process of HPBMCs activation induced by FIP-gts. On the other part, developing allergen of house dust mite Dermatophagoides pteronyssinus (Der p)-sensitized animal model, we evaluated whether FIP-gts could attenuate the symptoms of allergic asthma. The expression of IFN-γ in the splenocytes of Balb/c mice after feeding with FIP-gts for two weeks restimulated with FIP-gts was not increased but decreased instead. Using Differential Leucocyte Counting (DLC), we found that after sensitized with Der p increased the percentage of basophils in Balb/c mice blood, after fed with FIP-gts the basophils decreased to normal level. Unfortunately, we did not observe the percentage of eosinophils and total IgE were increased in asthma Balb/c mice. Our study demonstrated that FIP-gts regulated the proliferation of HPBMCs and induced the secretion of IFN-γ in HPBMCs via PI 3-kinase activation. Though we did not observed the symptoms of allergic asthma in Der p-sensitized mice, fed with FIP-gts could truly modify the immune response in Balb/c mice. It is suggested that FIP-gts may be as a health food with immunodulatory function.
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