ABSTRACT
The (1â6)-ß-D-glucans from Agaricus bisporus and Agaricus brasiliensis were purified to evaluate their effects on the innate immune system. THP-1 macrophages were used to investigate the induction of the expression of TNF-α, IL1ß, and COX-2 by RT-PCR. The purification of the polysaccharides gave rise to fractions containing 96-98% of glucose. The samples were analyzed by GC-MS, HPSEC and (13)C NMR, which confirmed the presence of homogeneous (1â6)-ß-D-glucans. The ß-glucans were incubated with THP-1 derived macrophages, for 3 h and 6 h to evaluate their effects on the expression of pro-inflammatory genes. Both ß-glucans stimulated the expression of such genes as much as the pro-inflammatory control (LPS). When the cells were incubated with LPS+ß-glucan, a significant inhibition of the expression of IL-1ß and COX-2 was observed for both treatments after 3 h of incubation. By the results, we conclude that the (1â6)-ß-D-glucans present an immunostimulatory activity when administered to THP-1 derived macrophages.
Subject(s)
Adjuvants, Immunologic/pharmacology , Agaricus/chemistry , Fungal Polysaccharides/pharmacology , Macrophages/immunology , beta-Glucans/pharmacology , Adjuvants, Immunologic/chemistry , Adjuvants, Immunologic/isolation & purification , Carbohydrate Conformation , Carbohydrate Sequence , Cell Differentiation , Cell Line , Cyclooxygenase 2/genetics , Cyclooxygenase 2/metabolism , Fungal Polysaccharides/chemistry , Fungal Polysaccharides/isolation & purification , Gene Expression/drug effects , Humans , Interleukin-1beta/genetics , Interleukin-1beta/metabolism , Lipopolysaccharides/pharmacology , Macrophages/drug effects , Molecular Sequence Data , Tumor Necrosis Factor-alpha/genetics , Tumor Necrosis Factor-alpha/metabolism , beta-Glucans/chemistry , beta-Glucans/isolation & purificationABSTRACT
The two main polysaccharides from the basidiomycetous fungus Laetiporus sulphureus were isolated, purified and characterized. The structural assignments were carried out using (13)C, (1)H, and (1)H,(13) HSQC nuclear magnetic resonance spectroscopy, methylation analysis, and Smith degradation. One was a linear beta-glucan having a (1-->3)-linked main chain, namely laminaran. The other was a fucomannogalactan, which consisted of a main chain of (1-->6)-linked alpha-D-galactopyranosyl residues, a part of them being substituted at O-2 by 3-O-D-mannopyranosyl-L-fucopyranosyl, alpha-D-mannopyranosyl and in a minor proportion, alpha-L-fucopyranosyl groups. This heteropolysaccharide is related to those of other Basidiomycetes heterogalactans, although it differs distinctly in its side-chain structures. Whereas part of the single-unit L-fucopyranosyl and/or 3-O-alpha-mannopyranosyl-L-fucopyranosyl residues are present as side chains of the other heterogalactans, additional alpha-D-mannopyranosyl units are present in our fucomannogalactan of L. sulphureus.
