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Mushrooms are known for their immune modulating effect for which the polysaccharide fraction, mainly glucans, seem to be responsible. Fungal β-glucans have been studied extensively, whereas little is known about mushroom α-glucans. We have earlier shown that the polysaccharide fraction from the mushroom A. bisporus, consisting 90% of α-glucans, induced in vitro tumor necrosis factor (TNF)α and nitric oxide production. The purpose of this study was to evaluate the effects of consuming.A. bisporus α-glucan on ex vivo cytokine production by human peripheral mononuclear blood cells (PBMCs). A double-blind randomized trial was designed in which 56 mildly hypercholesterolemic subjects consumed a control fruit juice with no added α-glucans (200 ml/day) for a 2-week run-in period. For the next 5 weeks, the control group (N=30) continued consumption of the control fruit juice, whereas the intervention group (N=26) consumed the same fruit juice enriched with α-glucans from A. bisporus (5 g glucans/day). Changes in interleukin (IL)-1β, IL-6 and TNFα cytokine production by lipopolysaccharide (LPS)-stimulated PBMCs were evaluated, as well as changes in T-helper (Th)1/Th2 cytokines by phytohemaggutinin (PHA)-stimulated PBMCs.Consumption of A. bisporus α-glucans lower LPS-induced TNFα production by 69% (P=0.017) as compared with the control group, whereas no effect on IL-1β and IL-6 was observed. No obvious Th1-Th2 skewing by PHA-stimulated PBMCs was observed. However, we observed a trend towards a decreased production of IL-12 and IL-10.Our current finding suggests that in vivo, α-glucans have lost their efficacy to stimulate the immune response as observed in our in vitro mouse model.