title: Effect of Agaricus brasiliensis-derived cold water extract on Toll-like receptor 2-dependent cytokine production in vitro.
abstract: Agaricus brasiliensis (Agaricus blazei Murrill) is well known as a medicinal mushroom. Fruit body of A. brasiliensis is rich in β-glucan and has shown benefits for various diseases. Both hot and cold water extraction are traditional methods for intake of this mushroom extract. In the present study, we prepared cold water extract of the fruit body of A. brasiliensis (ACWS). The 1,3-β-glucan segment of this fraction was too small and did not interact with the 1,3-β-glucan receptor, dectin-1. However, ACWS could induce production of various cytokines including IL-6 from murine splenocytes. Therefore, we aimed to identify the receptor that modulates IL-6 production using ACWS. We focused our attention on Toll-like receptors (TLRs) and examined them as follows. (i) The interaction between TLRs and ACWS was screened using HEK293 cells transfected with TLR plasmid. (ii) IL-6 production from splenocytes induced by ACWS was inhibited by treatment of anti-TLR antibodies. (iii) Direct binding activity between TLR protein and ACWS was assessed by ELISA-like assay. ACWS was found to activate HEK293 cells via TLR2, 4 and 5. However, only anti-TLR2 monoclonal antibody suppressed IL-6 production from splenocytes. In addition, ACWS has the ability to bind directly to TLR2 protein. Accordingly, we suggest that fruit body of A. brasiliensis has some water-soluble TLR ligand complexes, and TLR2 on splenocytes strongly induces IL-6 production.
authors: Laboratory for Immunopharmacology of Microbial Products, School of Pharmacy, Tokyo University of Pharmacy and Life Sciences, Horinouchi, Hachioji, Tokyo, Japan.
authors: Daisuke
authors: D
authors: Yamanaka
authors: Masuro
authors: M
authors: Motoi
authors: Ken-ichi
authors: K
authors: Ishibashi
authors: Noriko N
authors: NN
authors: Miura
authors: Yoshiyuki
authors: Y
authors: Adachi
authors: Naohito
authors: N
authors: Ohno
mesh_terms: D000364:Agaricus / Q000737:chemistry*; D000818:Animals; D045424:Complex Mixtures / Q000737:chemistry / Q000494:pharmacology; D005786:Gene Expression Regulation / Q000187:drug effects / Q000235:genetics; D057809:HEK293 Cells; D006801:Humans; D015850:Interleukin-6 / Q000096:biosynthesis* / Q000235:genetics; D037181:Lectins, C-Type / Q000819:agonists / Q000235:genetics / Q000378:metabolism; D008297:Male; D051379:Mice; D051195:Toll-Like Receptor 2 / Q000235:genetics / Q000378:metabolism*; D047071:beta-Glucans / Q000737:chemistry / Q000494:pharmacology; D000099181:Dectin-1
chemical_list: D045424:Complex Mixtures; D015850:Interleukin-6; D037181:Lectins, C-Type; D051195:Toll-Like Receptor 2; D047071:beta-Glucans; C546929:CLEC7A protein, human; D000099181:Dectin-1; C508600:IL6 protein, human; C495342:TLR2 protein, human; C493486:Tlr2 protein, mouse