Purification, Cloning, and Functional Characterization of a Novel Immunomodulatory Protein from Antrodia camphorata (Bitter Mushroom) That Exhibits TLR2-Dependent NF-kappa B Activation and M1 Polarization within Murine Macrophages

文化大學機構典藏 CCUR > 環境規劃暨生物資源學院 > 農學院 > 園藝暨生物技術學系 > 期刊論文 > Item 987654321/20548

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题名:	Purification, Cloning, and Functional Characterization of a Novel Immunomodulatory Protein from Antrodia camphorata (Bitter Mushroom) That Exhibits TLR2-Dependent NF-kappa B Activation and M1 Polarization within Murine Macrophages
作者:	Chien, PJ (Chien, Po-Jung) Sheu, F (Sheu, Fuu) Hsieh, KY (Hsieh, Kuang-Yang) Chin, KL (Chin, Kah-Lock) Huang, WT (Huang, Wan-Ting) Tsao, CY (Tsao, Chiao-Yin) Chen, YF (Chen, Yin-Fang) Cheng, HC (Cheng, Hui-Chung) Chang, HH (Chang, Hui-Hsin)
贡献者:	園生系
关键词:	Immunomodulatory protein Antrodia camphorata TLR2 NF-kappa B macrophage activation
日期:	2009
上传时间:	2011-11-30 15:27:43 (UTC+8)
摘要:	A new immunomodulatory protein, designated ACA, was purified from the mycelium extract of Antrodia camphorata, a well-known folk medicine bitter mushroom in Taiwan, and N-terminally sequenced. By taking advantage of its N-terminal amino acid sequence, the full-length ACA gene was cloned using rapid amplification of cDNA ends (RACE) approach. This gene encodes a 136 amino acid protein that is homologous to the phytotoxic proteins from fungi. On the basis of the data of N-terminal sequencing and N-glycosidase F treatment, the native ACA was confirmed to be a glycoprotein. The similarity in activation of TLR4-deficient macrophages by both the native ACA and recombinant ACA (rACA) suggested that the glycosyl group(s) of the native ACA was insignificant in macrophage activation. Moreover, the failure of rACA to induce TLR2-deficient macrophages and to activate the RAW 264.7 macrophages transfected with the dominate-negative MyD88 (dnMyD88) indicated that the ACA-mediated macrophage activation was TLR2/MyD88 dependent. Microarray assay of the ACA-activated NF kappa B-related gene expression showed that rACA demonstrated a LPS-mimetic proinflammatory response toward RAW 264.7 macrophages. Furthermore, rACA enhanced phagocytosis activity and CD86 (B7-2) expression as well as induced TNF-alpha and IL-1 beta production within murine peritoneal macrophages. A time-dependent induction of mRNA expression of cytokines TNF-alpha, IL-1 beta, IL-6, and IL-12 as well as chemokines CCL3, CCL4, CCL5, and CCL10, but not IL-10, CCL17, CCL22, and CCL24, was observed after the ACA treatment of the macrophages. These results proposed that ACA exhibited M1 polarization and differentiation in macrophages. Thus, ACA is an important immunomodulatory protein of A. camphorata.
显示于类别:	[園藝暨生物技術學系] 期刊論文

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