Mushroom (shiitake) extracts were dispersed with lecithin micelles to prepare superfine particles (0.05 to 0.2 μm in diameter) of β-1,3-glucan (micellary mushroom extracts). When mice were fed with these micelles of β-glucan (0.75 mg/day/mouse, smaller amounts of β-glucan), the number of lymphocytes yielded by the small intestine increased by up to 40%. More interestingly, the ratio of CD8αβ + TCRαβ + cells/CD8αα + TCRαβ + cells increased prominently. In parallel with this deviation in the distribution of lymphocyte subsets, tumor cytotoxicity against P815 cells and cytokine productions were also augmented. In other words, phylogenetically developed lymphocytes (CD8αβ + , TCRαβ + ) were much more effectively activated by the oral administration of micellary β-glucan. These results suggest that smaller amounts of micellary β-glucan might be useful for the potentiation of intestinal immunity.It is empirically known that mushrooms, especially components of β-glucan, are good for our health (2,6,10,14,17,21) and sometimes show anti-tumor effects in animal models and humans (3,4,15). However, such anti-tumor effects of crude mushroom extracts were limited in our preliminary experiments. To overcome this, we prepared superfine particles of β-1,3-glucan (i.e., mushroom extracts) dispersed with lecithin micelles using a high-pressure emulsifier in this study. These mushroom (shiitake) extracts were then used to examine whether oral administration has the potential to induce the augmentation of intestinal immunity in mice. Cumulative evidence has revealed that the augmentation of intestinal immunity is extremely important for immunological tolerance, anti-tumor effects, innate immunity against intracellular pathogens, etc. (1,7,8,13,18). The present results indicate that micellary mushroom extracts might have such potential via the augmentation of intestinal immunity, especially in the small intestine.
MATERIALS AND METHODSMice. C57BL/6 (B6) mice at the age of 8-10 weeks were used in this study. The mice were maintained in the animal facility of Niigata University (Niigata, Japan). All mice were fed under specific pathogenfree conditions. All experimental procedures were approved by the Committee on Animal Research of Niigata University.Oral administration of shiitake extracts. Shiitake mushrooms were broken down with a colloid mill and boiled at 95°C for 3 h in hot water, and the resulting extracts were filtered. These extracts were then mixed with lecithin and dispersed at 1500 kgf/ cm 2 with a high-pressure emulsifier (Ajinomoto Co. Inc., Kawasaki, Japan). Particles of β-1,3-glucan in the shiitake extracts (10~1000 μm) became smaller up to particle size of 0.05 to 0.2 μm (Fig. 1). The major component of the superfine particles was