Dual Effect of the Extract from the Fungus Coriolus versicolor on Lipopolysaccharide-Induced Cytokine Production in RAW 264.7 Macrophages Depending on the Lipopolysaccharide Concentration

Abstract

Extract from the fungus Coriolus versicolor (CV) is classified as an immunological response modifier. Previously, we have shown that this extract induces interleukin 6 (IL-6)-related extension of lipopolysaccharide (LPS)-induced fever. This study investigated the effect of CV extract on the production of pro-inflammatory cytokines and the expression of components of signal transduction pathways leading to the secretion of cytokines from RAW 264.7 macrophages stimulated with different doses of LPS. Methods: RAW 264.7 cells were stimulated with CV extract alone or co-treated with CV extract and LPS. The level of IL-6 and tumour necrosis factor α (TNF-α) in the culture media was measured using ELISA. Protein expression of Toll-like receptor (TLR) 4, phosphorylated IκB (p-IκB), CD14 glycoprotein and phospho-phosphatidylinositol 3-kinase (p-PI3K) was evaluated using Western blot. The effects of TLR4, nuclear factor κB (NF-κB) and p-PI3K on cytokine secretion were estimated using inhibitors: TAK-242, JSH-23 and LY294002. Results: CV extract itself stimulates the secretion of IL-6 and TNF-α and increases the expression of TLR4, p-IκB and p-PI3K. The presence of CV extract during the treatment of cells with lower concentrations of LPS (10 and 100 ng/mL) increases the cytokine production. Co-stimulation of cells with CV extract and LPS at a higher dose (500 ng/mL) decreases the secretion of cytokines. This effect is related to the changes in the expression of TLR4, CD14 glycoprotein, p-IκB and p-PI3K. Conclusion: This is the first report showing that the CV extract-induced production of cytokines is mediated by the PI3K signalling pathway. This extract acts antagonistically or additively with LPS on the production of IL-6 and TNF-α, depending on the LPS concentration. Our results are helpful for illustrating the mechanisms for the immunostimulatory effect of CV extract in inflammatory processes.

Endotoxin Tolerance Creates Favourable Conditions for Cancer Development

“…In our previous studies, we observed many times that macrophages are key cells involved in response to LPS [21,35]. It is commonly accepted that they may exhibit anti-cancer or pro-cancer properties, depending on their M1 and M2 phenotypes that are associated with the production of different cytokines.…”

“…An indicator of endotoxin tolerance (ET) is the decreased production of pro-inflammatory cytokines such as TNFα and IL-6 from macrophages during continued endotoxin exposure. Therefore, to determine the induction of ET, in this study, we evaluated the level of protein expression of TNFα and IL-6 in macrophages treated with a dose of 100 ng/mL of LPS based on the previous literature and preliminary data produced in our laboratory [21].…”

Exploring the Bioactive Mycocompounds (Fungal Compounds) of Selected Medicinal Mushrooms and Their Potentials against HPV Infection and Associated Cancer in Humans

“…Additionally, during treatment of cells with lower concentrations of lipopolysaccharide, the extract increased cytokine production, while higher dose of lipopolysaccharide led to their reduced synthesis. In other words, C. versicolor extract showed an antagonistic or additive effect according to lipopolysaccharide concentration [ 52 ].…”

Fever-Range Hyperthermia Promotes Macrophage Polarization towards Regulatory Phenotype M2b

“…RAW264.7 and J774A.1 cells were seeded in 24-well plates at the density of 5 × 10 5 cells/well. Following overnight pre-incubation and a 1 h pretreatment with TAK-242 at a concentration of 0.1 µM at 37 • C [44], 100 ng/mL of LPS was added, and cells were incubated for 2 h or 24 h (depending on the experiment) at 37 • C or 39 • C. After incubation, the cells were rinsed with ice-cold PBS and lysed in 100 µL of a 1 x RIPA buffer supplemented with 1% SDS and 0.5% protease inhibitor cocktail (all reagents were purchased from Sigma Aldrich). Following mechanical homogenization, the lysates underwent centrifugation to eliminate cellular debris.…”