Antibody-dependent cytotoxicity on chicken red blood cell targets mediated by the U937 cell line

• Published in: Immunology 1987-07, Vol.61 (3), p.277-282
• Main Authors: Villiers, M B, Ward, R H, Lachmann, P J
• Format: Article
• Language: English
• Subjects: Animals; Antibody-Dependent Cell Cytotoxicity - drug effects; Cell Line; Chickens; Cytoskeleton - drug effects; Erythrocytes - immunology; Humans; Iron - metabolism; Lymphoma, Large B-Cell, Diffuse - immunology; Oxygen - metabolism; Protein Biosynthesis; Tetradecanoylphorbol Acetate - pharmacology; Trypsin Inhibitors - pharmacology
• ISSN: 0019-2805; 1365-2567

We have characterized a model system for the study of antibody-dependent cytotoxicity (ADCC) mediated by human macrophages and monocytes. The U937 cell line is used as a source of effector cells. We confirmed a previous report (Gidlund et al., 1981) that U937 can be activated using PMA to kill in ADCC, and the characteristics of the observed cytotoxicity are described. Activation of effectors was maximal after 20-hr preincubation in the presence of 10 ng/ml PMA. In these conditions, lysis was approximately 70% in 2 hr at an effector to target ratio of 5:1. Activation correlated with the expression of complement receptors CR1 and CR3. No antibody-independent cytotoxicity was observed. The effects of various inhibitors of oxygen species were investigated: the lysis obtained in the above conditions was inhibited at 50% in the presence of either 450 mM dimethyl sulphoxide or 30,000 U/ml catalase, but superoxide dismutase (up to 10,000 U/ml) or ferricytochrome c (up to 2 mM) had no effect. The same inhibition was observed with 40 mM desferrioxamine or with 1 mM 0-phenanthroline, which are both iron scavengers, or in the presence of 300 microM colchicine or 1.5 microM dihydrocytochalasin B, which are two inhibitors of cytoskeletal functions. An identical effect was obtained in the presence of 1 TIU/ml bovine pancreas trypsin inhibitor, whereas soya bean trypsin inhibitor, which is more specific, had no effect up to 5000 BAEE U/ml. No inhibition was seen with protein synthesis inhibitors as cycloheximide or puromycin at 40 micrograms/ml. The significance of these results is discussed.