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Technique for screening immune-enhancing polysaccharides in food using 1,25-dihydroxyvitamin D3-differentiated HL60 cells.

Monobe M, Ema K, Kato F, Hirokane H, Maeda-Yamamoto M

National Institute of Vegetable and Tea Science, 2769 Kanaya, Shimada, Shizuoka 428-8501, Japan. monobe@affrc.go.jp

A technique for screening immune-enhancing polysaccharides in food using the phagocytotic activity of 1,25-dihydroxyvitamin D3 (VD3)-differentiated HL60 cells is presented. HL60 cells, a human acute promyelocytic cell line, can differentiate along the monocytic lineage following exposure to VD3 or phorbol-12-myristate-13-acetate (PMA). For differentiated cells along the monocytic pathway, HL60 cells were maintained in RPMI 1640 medium supplemented with 10% FBS and 120 nM VD3 for more than 1 week. VD3-differentiated HL60 cells were seeded into 48-well plates, YG-labeled microspheres and polysaccharides were added and mixed using a plate shaker at 1100 rpm for 30 s, and then the mixture was incubated overnight at 37 degrees C in 5% CO2. The cells were fixed with 2% formaldehyde and resuspended in phosphate-buffered saline. The rate of phagocytosis was measured with a flow cytometer. VD3-differentiated cells but not non- and PMA-differentiated cells resulted in an elevation of phagocytic activity by various immune-enhancing polysaccharides in foods.

Published 28 March 2007 in J Agric Food Chem, 55(7): 2543-7.
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