Augmentation of monocyte-mediated cytocidal activity by a low dose tumour necrosis factor measured by the kinetic colorimetric microplate assay.
This paper describes a simple kinetic colorimetric assay for the quantitation of human peripheral blood monocyte-mediated cytotoxic activity against tumour cells. Isolated effector monocytes were cultured overnight with an increasing number of target cells in 96-well microplates. Cytotoxic activity of monocytes was determined by modified nitroblue tetrazolium (MTT) dye assay using standard ELISA reader offering possible automation. The test was performed with three different effector/target cell ratios using a fixed number of monocytes. This allowed the expression of cytotoxic activity of monocytes in cytotoxic activity units. The assay was found to be a simple method to demonstrate that low doses of TNF (1 U/ml) enhanced monocyte-mediated cytotoxicity.
Siedlar
M
M
Department of Clinical Immunology, Copernicus Medical School, Cracow, Poland.
Uracz
W
W
Zembala
M
M
D003124:Colorimetry; D003601:Cytotoxicity Tests, Immunologic / Q000379:methods*; D003602:Cytotoxicity, Immunologic; D006801:Humans; D007700:Kinetics; D008433:Mathematics; D008546:Melanoma, Experimental / Q000276:immunology; D009000:Monocytes / Q000276:immunology*; D012044:Regression Analysis; D013778:Tetrazolium Salts; D013844:Thiazoles; D014407:Tumor Cells, Cultured; D014409:Tumor Necrosis Factor-alpha / Q000276:immunology*
D013778:Tetrazolium Salts; D013844:Thiazoles; D014409:Tumor Necrosis Factor-alpha; C022616:thiazolyl blue