Technique for Screening Immune-Enhancing Polysaccharides in Food Using 1,25-Dihydroxyvitamin D3-Differentiated HL60 Cells

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 °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.

Keywords: HL60; 1,25-dihydroxyvitamin D3; immune-enhancing polysaccharides; phagocytosis