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Nutritional Immunology

Sodium Cromoglycate Inhibits Absorption of the Major Soybean Allergen, Gly m Bd 30K, in Mice and Human Intestinal Caco-2 Cells¹

² Laboratory of Molecular Function of Food, Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Uji, Kyoto 611-0011, Japan; ³ Department of Food Science, Graduate School of Nutrition and Bioscience, The University of Tokushima, Tokushima 770-8503, Japan; ⁴ Department of Applied Cell Biology, Graduate School of Agriculture, Kinki University, Nara 631-8505, Japan; and ⁵ Department of Nutritional Science for Well-Being, Faculty of Health Science for Welfare, Kansai University of Welfare Science, Kashihara, Osaka 582-0026, Japan

^* To whom correspondence should be addressed. E-mail: fat{at}kais.kyoto-u.ac.jp.

Our previous data showed that Gly m Bd 30K was absorbed from the gastrointestinal tract and circulated in blood in mice. This study was conducted to determine the mechanism and identify the inhibitor of such absorption. Using sandwich ELISA and immunoblotting, we found that intact Gly m Bd 30K was absorbed from apical to basolateral solutions and intracellularly accumulated by Caco-2 cells in a dose- and time-dependent manner. The absorption and intracellular accumulation of Gly m Bd 30K were significantly suppressed when Caco-2 cells were treated with sodium cromoglycate (SCG) (0-50 mmol/L) in a dose-dependent manner. In 24-d-old mice orally treated with SCG (10–1000 mg/kg body weight), plasma Gly m Bd 30K concentration decreased significantly 30–120 min after Gly m Bd 30K (2000 mg/kg body weight) administration. Moreover, inhibitors that suppress the clathrin-dependent endocytosis dansylcadaverine, the caveolae-dependent endocytosis nystatin and clathrin, and the caveolae-dependent endocytosis methyl-ß-cyclodextrin had inhibitory effects on the absorption and intracellular accumulation of Gly m Bd 30K by Caco-2 cells. These data indicate that Gly m Bd 30K is absorbed and intracellularly accumulated in Caco-2 cells via clathrin- or caveolae-dependent endocytosis. We propose that the absorption and intracellular accumulation of Gly m Bd 30K are inhibited by SCG via clathrin- or caveolae-dependent endocytosis.