本研究以遺傳工程的方式表現松杉靈芝免疫調節蛋白質，純化得FIP-gts蛋白。在In vitro 實驗中證實松杉靈芝免疫調節蛋白質對於人類周邊血細胞cytokine的產量其活性相當，處理2 ug/ml 的FIP-gts時，經 24 小時及48小時後會誘發產生大量IFN-r 高達4135 pg/ml。靈芝免疫調節功能蛋白質透過磷酸化的訊息傳遞促進增加 IFN-γ的表現，因此靈芝可藉著調節磷酸化的狀況，特別以LY294002的抑制劑的抑制效果特別明顯。所以是透過PI-3 kinase的途徑來表現其誘發IFN-r的能力。而也觀察到靈芝免疫調節功能蛋白質顯著的誘發磷酸化的Erk蛋白表現增加可能與此蛋白會促進T細胞增殖有密切關係。以8-10周大BalB/C Male老鼠進行餵食試驗，經由每二天餵食FIP-gts (50 ug)共餵食7次其spleen 細胞與餵食水的spleen細胞數目由3-5 X 107劇增到 1-2 X 108細胞數，所以確實可增加老鼠脾臟細胞增加，應該具有免疫調節功能但細胞激素的測量仍持續進行中。且分析確實都有降低由塵蹣所誘發的eosinophil數目，但唯一目前美中不足的地方是過敏老鼠並無明顯觀察得到老鼠會氣喘且Neutrophils隨濃度增加也會增加。所以本計劃主要目的靈芝免疫調節功能蛋白對於IFN-r表現量做為標的，已可當作初步評估靈芝免疫調節功能蛋白的免疫調節活性功能預防由塵璊(Derp)所引起的氣喘。但若發病後的治療方面仍有待評估。 In this study we expressed and purified the fungal immunomodulatory protein from Ganoderma tsugae by molecular cloning. It is designate FIP-gts. The effects of FIP-gts induced on human T cell activations and secreted the significant production of IFN-r. By analyzing the cytokine, FIP-gts were shown to act on T cells type I by stimulating them to secrete a large amount of IFN-r till to the concentration of 4315 pg/ml for 48 hr. Fungal immunomodulatory protein induce the production of Interferon-r through the phosphorylation signal transduction pathway. Specific PI-3 kinase inhibitor, Ly294002 could abolish the production of IFN-r by FIP-gts. It may induce the production of IFN-r through PI-3 kinas activation. Simultaneously, FIP-gts also stimulate the phosphorylation of ErK due to stimulation of proliferation of T cells. The numbers of spleen cells will increase from 3 X107 to 1 X108 cells by feeding the FIP-gts. It is very exciting that FIP-gts could decrease the numbers of eosinophil induced by Derp. It is illustrate that FIP-gts is a good food for preventing from asthma. Unfortunately, it also increase the numbers of neutrophils dependent on the concentration of FIP-fve. In the further, we will study the therapy of asthma.
