在本計劃中我們分析了鴻喜菇、鮑魚菇、杏鮑菇、柳松菇、袖針菇及金針菇等食用菇蛋白質，發現柳松菇蛋白質會促進人類周邊淋巴球細胞的TNF-a分泌，而由金針菇真菌類免疫調節蛋白FIP-fve (Fungal Immunomodulatory Protein Flammulina velutipes)當中，利用PI染色與流式細胞儀分析，發現FIP-fve可促使HPBMCs (Human peripheral blood mononuclear cells)的細胞週期由G0/G1期進入S期進而造成細胞增生，且大量誘發IFN-r及TNF-a等Th1的細胞激素，對於interleukin-4只有稍許誘導。在此研究中我們利用重組蛋白工程在大腸桿菌中表現並純化得reFIP-fve。而預先處理MAPK的pp38專一性抑制劑SB203580則只可部份抑制由FIP-fve所誘導之S期的增加。另一方面分析FIP-fve誘導HPBMCs產生的細胞激素時，發現IFN-γ持續大量表現，所以此類FIP-fve是趨向活化Th1細胞；而預先處理SB203580則可完全抑制FIP-fve誘導之IFN-γ的表現；此外以Western blot方式證實，以FIP-fve處理HPBMCs後會磷酸化而活化pp38的下游基因，由此顯示MAPK(p38)在FIP-fve活化HPBMCs的過程中扮演一個相當重要的角色。FIP-fve應非常有潛力發展成有效預防及治療由Th2所誘發的過敏反應。 FIP-fve is a protein purified from Flammulina velutipes, an edible golden needle mushroom thought to possess potent immunomodulatory properties. When examined for its effects on lymphocytes, FIP-fve exhibited potent mitogenic effects on human peripheral blood lymphocytes, inducing G1/G0 to S phase proliferation. T cells activated by FIP-fve show significant production and secretion of interferon-r but low detectable levels of IL-4 in vitro or in vivo. In addition, activation of hPBLs with FIP-fve results in increased production of IFN-r associated with ICAM-1 expression and p38 MAPK kinase activation. SB203580 can fully abolish the production of IFN-r induced by FIP-fve. At the same time, SB203580 only partially prevents the lymphocytes from progressing from G1 to S phase of the cell cycle. These findings demonstrate that FIP-fve is a potent T cell activator, mediating its effects via cytokine regulation of MAPK kinase. The immunoprophylatic effects of FIP-fve in Th2-mediated allergic anaphylaxis are believed to be associated with the ability of FIP-fve to enhance activation of IFN-r releasing Th1 cells.
