Chen, Chia-JungChia-JungChenKrishna, R.VijayaR.VijayaKrishnaTsai, Chia-CheChia-CheTsaiWu, Wan-HsunWan-HsunWuChao, Louis KuopingLouis KuopingChaoHwang, Kent-HaoKent-HaoHwangChien, Chichen MichaelChichen MichaelChienChang, Hwan-YouHwan-YouChangChen, Shui-TeinShui-TeinChen2012-10-202018-07-062012-10-202018-07-062010http://ntur.lib.ntu.edu.tw//handle/246246/243463The preserved fungal species Antrodia camphorata has diverse health-promoting effects and has been popularly used in East Asia as a traditional herb. We isolated a volatile compound from the culture medium of A. camphorata and identified it as γ-dodecalactone (γ-DDL). Cytomic screening for immune-modulating activity revealed that γ-DDL can activate human NK cells to express the early activation marker CD69. Further experiments showed that γ-DDL not only can induce NK cells to express CD69 but also stimulate NK cells to secrete cytotoxic molecules (FasL and granzyme B) and Th1 cytokines (TNF-α and INF-γ). Measuring the distribution of γ-DDL in the subcellular compartments of NK cells revealed that γ-DDL has been converted to 4-hydroxydodecanoic acid (an acyclic isomer of γ-DDL) in a time-dependent manner in the cytoplasm. Synthetic (R,S)-4-hydroxydodecanoic acid activated NK cells to express CD69 mRNA within 10 min, in contrast to γ-DDL, which activated NK cells to express CD69 within 50 min. This faster activation suggests that γ-DDL has converted to 4-hydroxydodecanoic acid and to stimulate the NK cells to express CD69. Optically pure (R)-(+)-4-hydroxydodecanoic acid and (S)-(-)-4-hydroxydodecanoic acid were obtained via: (1) synthesis of its diastereomeric esters of (R,S)-4-hydroxydodecanoic (R)-(-)-2-phenylpropionate; (2) separation of diastereomers via preparative HPLC, and (3) subsequent hydrolysis of the obtained optical pure ester of (R)-(+)-4-hydroxydodecanoic acid (R)-(-)-2-phenylpropionate and (R)-(-)-4-hydroxydodecanoic acid (R)-(-)-2-phenylpropionate, respectively. Further assays of NK cells activation using each enantiomer showed that only the (R)-(+)-4-hydroxydodecanoic acid can activate NK cells. ? 2010 Elsevier Ltd. All rights reserved.en-USγ-Dodecalactone; 4-Hydroxydodecanoic acid; Antrodia camphorata; CD69; NK cells[SDGs]SDG32 phenylpropionate; 4 hydroxydodecanoic acid; CD69 antigen; Fas ligand; fungal protein; gamma dodecalactone; gamma interferon; granzyme B; immunomodulating agent; tumor necrosis factor alpha; unclassified drug; Antrodia camphorata; article; cell activation; cell compartmentalization; concentration response; controlled study; cytoplasm; cytotoxicity; diastereoisomer; drug identification; drug isolation; enantiomer; fungus culture; high performance liquid chromatography; human; human tissue; hydrolysis; natural killer cell; protein expression; structure activity relation; 4-Butyrolactone; Antigens, CD; Antigens, Differentiation, T-Lymphocyte; Antrodia; Fas Ligand Protein; Granzymes; Humans; Interferon-gamma; Killer Cells, Natural; Lectins, C-Type; Lymphocyte Activation; Stereoisomerism; Tumor Necrosis Factor-alpha; Taiwanofungus camphoratusjournal article10.1016/j.bmc.2010.07.024http://ntur.lib.ntu.edu.tw/bitstream/246246/243463/-1/115.pdf