自一些真蕈類中(金針菇、草菇、靈芝、洋菇、猴頭菇及杏鮑菇)分離得凝血活性部分及溶血活性部分中篩選出具有免疫調節功能活性蛋白。靈芝免疫調節功能蛋白質會促進ICAM-1 (Intercellular adhesion molecule-1)的表現，並明顯增加IFN-γ，因此靈芝增加CD2表現，所以靈芝可藉著調節Immunocompetent cell上Adhesion分子來表現其藥性。結果發現松杉靈芝、草菇及金針菇中可分離純化得此類免疫調節功能蛋白質。分別自松杉靈芝子實體及菌絲體中分別純化得14 kDa大小的蛋白質，竟意外發現只有菌絲體具有此免疫調節活性蛋白將之命名為FIP-gts。而靈芝屬菌株中，依許瑞祥博士論文中分離區別鑑定G. applanatum、G. formosanum、G. fornicatum、G. lucidium、G. tropicum及G. tsugae等數種靈芝。每種菌種間其免疫調節活性差異相當大，目前G. lucidium及G. tsugae已被證實具有免疫調節功能，但應用於氣喘預防方面尚未被證實。況且其他菌種如本計畫材料Ganoderma tsugae(FIP-gts)仍有待證實。本研究並將此蛋白的基因選殖並以大腸桿菌表達系統表現此蛋白。而FIP-gts此免疫調節功能是以刺激人類週邊血淋巴球細胞所誘發產生Interfero-r能力及誘導BALB/C老鼠淋巴細胞誘導Interferon-r能力作為判斷，使用Dyna-bead細胞分離系統分離Non-B與Non-T細胞，發現FIP-gts只有在Non-B細胞群中具有細胞增生能力。塵璊(DerpII)由於蔡考圓教授已到新加坡大學任教，曾與其通過E-mail，提及不適合提供DerpII表現載體，但願意考慮直接提供蛋白，由於動物實驗所以所需蛋白非常多，所以擬自己製備，以另二種管道獲得。所以另一由國泰醫院免疫風濕科主任蔡肇基醫師提供DerpII表現載體；並得其同意，所以正進行製備純化。並以其所建立的氣喘性老鼠為實驗模式。 A fungal immunomodulatory protein (FIP-gts) has been isolated and purified from the Ganoderma tsugae mushroom. The apparent molecular mass of FIP-gts was determined by SDS/PAGE with 13 kDa. The immunomodulatory activity of FIP-gts was demonstrated by its stimulatory activity toward human peripheral blood lymphocytes and remarkable induction of interferon-gamma (IFN-r) cytokine. It may play an role on stimulating helper T1 cell pathway. We have observed the high similarity between FIP-gts and another immunomodulatory protein, FIP-fve, isolated from Flammulina velutipes. FIP-fve have shown to suppress systemic anaphylaxis reactions and local swelling of mouse edema by compound 48/80. Ganoderma, an oriental fungus, has been widely used as a remedy for promotion of health and lingevity. It was reported to be effective in the treatment of chronic hepatopathy and anti-tumor activity. But, the immunomodulatory activity of FIP-gts will be further clarified. In asthma patients, it is characterized by skewed polarization of helper T cell (Th) development toward T helper cells type 2 (Th2) with increased production of interleukin 4 (IL-4) and IL-5, which affect immunoglobulin (Ig) isotype switching toward IgE, and differentiation, growth of eosinophils, respectively. Immune responses to protein antigens are strongly influenced by the nature of the helper T lymphocyte (Th) subsets participating in the response. Once deviation toward a Th1 or Th2 response begins, there is potential for further polarization driven by the Th cells themselves. The aim of the present study will to examine the in vivo effect of FIP-gts on a murine model of asthma induced by Dermatophagoides pteronyssinus derived DerpII allergen. BALB/C mice immunized with DerpII.
