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ORIGINAL RESEARCH ARTICLE

Activation of Selective Transcription Factors and Cytokines by Water-Soluble Extract from Lentinus lepideus

Mirim Jin^*, Hyung Jin Jung^*, Jeong June Choi, Hyang Jeon^*, Jin Hwan Oh^*, Bongcheol Kim^*, Sung Seup Shin^*, Jong Kyu Lee, Keejung Yoon and Sunyoung Kim^*,,1

^* PanGenomics Co. Ltd., Biotechnology Incubating Center, Seoul 151-742, Korea;
Tree Pathology and Mycology Laboratory, Kangwon National University, Chunchon, 200-701, Korea;
School of Biological Science and Institute of Molecular Biology and Genetics, Seoul National University, Seoul 151-742, Korea

We isolated a water-soluble extract, PG101, from cultured mycelia of Lentinus lepideus. Treatment of human peripheral blood mononuclear cells (PBMCs) with PG101 increased levels of TNF-, IL-1ß, IL-10, and IL-12 by 100- to 1000-fold, whereas GM-CSF and IL-18 were activated by an order of magnitude. On the contrary, IFN- and IL-4 were not affected. The response to PG101 occurred in a dose- and time-dependent manner. From the human PBMCs treated with PG101, TNF- was a first cytokine to be activated, detectable at 2 hr post-treatment followed by IL-1ß at 6 hr post-treatment. IL-12 and IL-10 were the next to follow. GM-CSF and IL-18 both showed significant increases 24 hr after treatment. When PBMCs were sorted into various cell types, monocyte/macrophages, but not T and B cells, were the major target cell type responsive to PG101. Consistent with this result, the profile of cytokine expression upon PG101 treatment was comparable between PBMCs and a human promonocytic cell line (U937), whereas cell lines of T cell and myeloid origins did not respond to PG101. Data from a transient transfection assay involving specific reporter plasmids indicated that cellular transcription factor such as NF-B, but not AP-1, was highly activated by PG101. Results from a gel retardation assay and the experiment involving a specific NF-B inhibitor confirmed the involvement of NF-B. Despite its significant biological effect on various cytokines, PG101 remained nontoxic in both rats and PBMCs even at a biological concentration approximately 20 times greater. PG101 demonstrates great potential as a therapeutic immune modulator.

Key Words: Lentinus lepideus • PG101 • immune modulator • monocyte/macrophage • cytokines • transcription factor • NF-B

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