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 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.
Monobe
Manami
M
National Institute of Vegetable and Tea Science, 2769 Kanaya, Shimada, Shizuoka 428-8501, Japan. monobe@affrc.go.jp
Ema
Kaori
K
Kato
Fumiko
F
Hirokane
Hisako
H
Maeda-Yamamoto
Mari
M
D002117:Calcitriol / Q000494:pharmacology*; D002454:Cell Differentiation / Q000187:drug effects*; D005504:Food Analysis; D018922:HL-60 Cells; D006801:Humans; D007109:Immunity / Q000187:drug effects*; D010587:Phagocytosis / Q000187:drug effects; D011134:Polysaccharides / Q000032:analysis* / Q000494:pharmacology*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
D011134:Polysaccharides; D002117:Calcitriol; D013755:Tetradecanoylphorbol Acetate