真菌類免疫調節功能蛋白質（Fungal immunomodulatory proteins, FIPs）可以刺激T淋巴球細胞增殖，減低Compound 48/80發炎反應，且會促進IFN-r的大量表現。本計劃主要發現自金針菇純化所得的FIP不論在老鼠spleen，或人類周邊血細胞皆誘發大量IFN-r表現，並同時也會誘發interferon-r所調控的NO產生。且研究FIPs是經何種途徑誘導IFN-r產生，結果FIP可能是間接透過p38活化，進而結合到IFN-r promoter區位，所以將以Western blot偵測FIPs會增加p38蛋白的6倍表現量影響。好像是透過p38的表現已Western blot證實誘發p38產生。本計劃並自CD4+ T細胞株中，構築IFN-r promoter（543 bp）並具有螢光（Luciferase）報告基因的質體，藉由網站預估分析IFN-r promoter的Transcription factor binding site進而轉染過的細胞中Luciferase活性，來了解FIPs與IFN-r promoter調控關係，並知道其Promoter區位的序列何者重要。 The immunomodulatory activity of FIPs were demonstrated by their stimulatory activity toward human peripheral blood lymphocytes, and its anti-inflammatory of compound 48/80 stimulated edema. FIPs were found to enhance the transcriptional expression and translation expression of Interferon-gamma and Interleukine-2. FIPs can induce the IFN-r expression toward human peripheral blood mononuclear cells (PBMCs) and mice spleen cells. FIPs also induced the production of NO from the downstream of interferon-r regulated. Inducible gene expression in eukaryotes is mainly controlled by the activity of transcriptional activator proteins, such as p38. We will investigate the 6 fold induction of p38 Western blot. IFN-r promoter (543 bp) was ligated into pGL-3 luciferase reporter gene. In the present study, we will characterize a regulatory element in the human interferon-r promoter by FIPs. Transient transfection of IFN-r promoter-reporter gene construct in the human T cell line Jurkat cells will exhibit that FIPs stimulated the transcription of IFN-gamma promoter driven luciferase gene.
