Augmentation of monocyte-mediated cytocidal activity by a low dose tumour necrosis factor measured by the kinetic colorimetric microplate assay

Immunol Lett. 1992 Dec;34(3):249-56. doi: 10.1016/0165-2478(92)90221-9.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Colorimetry
  • Cytotoxicity Tests, Immunologic / methods*
  • Cytotoxicity, Immunologic
  • Humans
  • Kinetics
  • Mathematics
  • Melanoma, Experimental / immunology
  • Monocytes / immunology*
  • Regression Analysis
  • Tetrazolium Salts
  • Thiazoles
  • Tumor Cells, Cultured
  • Tumor Necrosis Factor-alpha / immunology*

Substances

  • Tetrazolium Salts
  • Thiazoles
  • Tumor Necrosis Factor-alpha
  • thiazolyl blue