title: Generic tools to assess genuine carbohydrate specific effects on in vitro immune modulation exemplified by β-glucans.
abstract: Even if carbohydrate preparations from plant/fungal sources have a high degree of purity, observed immune-stimulation may be caused by minute sample contaminations. Using the example of different β-glucans we present a range of analytical tools crucial for validation of possible immune-stimulatory effects. Two yeast (MacroGard and Zymosan) and one cereal β-glucan (CBG40) increased IL-8 secretion by HT-29 cells considerably. Degradation of the β-glucan samples with β-glucan specific enzymes did hardly influence the effect of Zymosan and CBG40 but significantly decreased the effect of MacroGard. Stimulation of IL-8 secretion by CBG40 and Zymosan was hence not due to their β-glucan content. Instead, the effect of the CBG40 sample was due to low levels of LPS despite the inability of the known LPS inhibitor Polymyxin B to supress its stimulatory effect. We conclude that targeted enzymatic degradation of samples is a powerful validation tool to investigate carbohydrate specific immune-modulation.
authors: Nofima, Norwegian Institute of Food, Fishery and Aquaculture Research, PB 210, N-1431 Aas, Norway. anne.rieder@nofima.no
authors: Anne
authors: A
authors: Rieder
authors: Stine
authors: S
authors: Grimmer
authors: Finn L
authors: FL
authors: Aachmann
authors: Bjørge
authors: B
authors: Westereng
authors: Svein Olav
authors: SO
authors: Kolset
authors: Svein Halvor
authors: SH
authors: Knutsen
mesh_terms: D000276:Adjuvants, Immunologic / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology*; D002523:Edible Grain / Q000737:chemistry; D004731:Endotoxins / Q000032:analysis; D006026:Glycoside Hydrolases / Q000378:metabolism; D019073:HT29 Cells; D006801:Humans; D006868:Hydrolysis; D016209:Interleukin-8 / Q000378:metabolism; D011786:Quality Control; D015203:Reproducibility of Results; D012441:Saccharomyces cerevisiae / Q000737:chemistry; D047071:beta-Glucans / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology*
chemical_list: D000276:Adjuvants, Immunologic; D004731:Endotoxins; D016209:Interleukin-8; D047071:beta-Glucans; D006026:Glycoside Hydrolases