Disruption of Actin Cytoskeleton Enhanced Cytokine Synthesis of Splenocytes Stimulated with Beta-Glucan from the Cauliflower Medicinal Mushroom, Sparassis crispa Wulf.:Fr. (Higher Basidiomycetes) in Vitro

Abstract

Beta-glucan (BG) is a representative pathogen-associated molecular pattern (PAMP) produced by pathogenic fungi. SCG is a BG obtained from Sparassis crispa, which stimulates splenocytes in DBA/2 mice to produce cytokines, such as GM-CSF, IFN-γ, and TNF-α. In the present study, we analyzed the molecular mechanism of SCG-mediated cytokine synthesis using cytocharasin D (CytD), an inhibitor of actin polymerization. It was found that GM-CSF and TNF-α synthesis of splenocytes stimulated with SCG, but not with lipopolysaccharide, was significantly enhanced in the presence of CytD. CRDO, partially hydrolyzed linear 1,3-BG curdlan, stimulated splenocytes of DBA/2 mice slightly to produce cytokines. CRDO, acting as an antagonist in the presence of SCG, changed to a strong agonist in the presence of CytD. CytD also enhanced cytokine synthesis of bone marrow-derived dendritic cells. Taken together, cytokine productivity of BG was significantly dependent on molecular weight, and CytD treatment is useful to enhance the sensitivity for analyzing the immunostimulating activity of BG in vitro.

Beta-glucans from edible and medicinal mushrooms: Characteristics, physicochemical and biological activities

“…In another study, the dried mushroom Flammulina velutipes was milled and submitted to aqueous and alkaline extractions, and then the crude b-glucan residue was subjected to further alkaline extraction at 100 8C, using 2% and then 25% KOH (Smiderle et al, 2006). b-Glucan from mushroom Sparassis crispa was prepared using cold alkali (10% NaOH/5% urea) at 4 8C with a 2-day extraction time, and subsequently the extract was applied to a DEAE Sephadex A25 column for further purification (Shibata et al, 2012). Sen et al (2013) also used distilled water (100 8C) to extract b-glucan from a hybrid mushroom, pfls1h of P. florida and L. squarrosulus (Mont.)…”

Natural Products and Biological Activity of the Pharmacologically Active Cauliflower MushroomSparassis crispa

“…Shibata et al found that both GM-CSF and TNF- α synthesis in DBA/2 mouse splenocytes stimulated with SCG, but not with lipopolysaccharide, were significantly enhanced in the presence of cytochalasin D (CytD), an inhibitor of actin polymerization [27]. On the other hand, Kim et al pointed out the importance of the role of Toll-like receptor 4 (TLR4) [28].…”

Sparassis crispa Intake Improves the Reduced Lipopolysaccharide-Induced TNF-α Production That Occurs upon Exhaustive Exercise in Mice

“…Sparassis crispa is an edible mushroom that contains high amounts of β-glucan, more than 30% of the dry weight of the fruiting bodies, compared with levels in other mushrooms [6]. Several studies have revealed that consumption of S. crispa and β-glucan extracted from S. crispa has immunopotentiative effects, such as enhanced inflammatory cytokine production in splenocytes, improved peripheral NK cell activity, and anti-tumor action [6,7,8,9]. Based on these results, S. crispa consumption enhances immune function.…”