title: The TLR2 agonist in polysaccharide-K is a structurally distinct lipid which acts synergistically with the protein-bound β-glucan.
abstract: Protein-bound polysaccharide-K (Krestin; PSK) is a hot-water extract of Trametes versicolor with immune stimulatory activity. It has been used for the past 30 years and has demonstrated anti-tumor efficacy in multiple types of cancer. The ability of PSK to activate dendritic cells and T cells is dependent on its ability to stimulate Toll-like receptor 2 (TLR2), yet it remains unknown which structural component within PSK activates TLR2. The purpose of this study was to identify the TLR2 agonist within PSK and understand its role in the overall mechanism of PSK's immunogenic activity. TLR2 activity was eliminated by treatment with lipoprotein lipase but not by trypsin or lyticase. Rapid centrifugation of PSK can separate the fraction with TLR2 agonist activity from the soluble β-glucan fraction. To study the potential interaction between the β-glucan component and the lipid component, we labeled the soluble β-glucan with fluorescein. Uptake of the labeled β-glucan by J774A macrophages and JAWSII dendritic cells was inhibited by anti-Dectin-1 antibody but not by anti-TLR2 antibody, confirming that Dectin-1 is the receptor for β-glucan. Interestingly, pre-treatment of JAWSII cells with the TLR2-active lipid fraction significantly enhanced the uptake of the soluble β-glucan, indicating the synergy between the TLR2 agonist component and the β-glucan component. Altogether, these results present evidence that PSK has two active components-the well-characterized protein-bound β-glucan and a previously unreported lipid-which work synergistically via the Dectin-1 and TLR2 receptors.
authors.affiliation: Tumor Vaccine Group, Department of Medicine, University of Washington, 850 Republican Street, Seattle, WA, 98109, USA, quaylek@uw.edu.
authors.forename: Kenneth
authors.initials: K
authors.lastname: Quayle
authors.forename: Catherine
authors.initials: C
authors.lastname: Coy
authors.forename: Leanna
authors.initials: L
authors.lastname: Standish
authors.forename: Hailing
authors.initials: H
authors.lastname: Lu
mesh_terms: D000276:Adjuvants, Immunologic / Q000737:chemistry / Q000494:pharmacology; D000818:Animals; D000970:Antineoplastic Agents / Q000737:chemistry / Q000494:pharmacology; D001688:Biological Products / Q000737:chemistry / Q000494:pharmacology; D003713:Dendritic Cells / Q000187:drug effects* / Q000378:metabolism; D004357:Drug Synergism; D006801:Humans; D037181:Lectins, C-Type / Q000378:metabolism; D008055:Lipids / Q000494:pharmacology*; D008264:Macrophages / Q000187:drug effects* / Q000378:metabolism; D051379:Mice; D011509:Proteoglycans / Q000737:chemistry / Q000494:pharmacology*; D013601:T-Lymphocytes / Q000187:drug effects / Q000378:metabolism; D051195:Toll-Like Receptor 2 / Q000819:agonists*; D055454:Trametes / Q000737:chemistry*; D047071:beta-Glucans / Q000494:pharmacology*; D000099181:Dectin-1
chemical_list: D000276:Adjuvants, Immunologic; D000970:Antineoplastic Agents; D001688:Biological Products; D037181:Lectins, C-Type; D008055:Lipids; D011509:Proteoglycans; D051195:Toll-Like Receptor 2; D047071:beta-Glucans; D000099181:Dectin-1; C010770:polysaccharide-K