78 - ANTIBODY-DEPENDENT CELLULAR CYTOTOXICITY (ADCC) OF ERYTHROID AND TUMOR CELLS

Abstract

Publisher Summary This chapter describes and discusses a short-term 51Cr-release ADCC assay using purified mononuclear phagocytes as effector cells against hapten-modified antibody-coated target cells. The macrophages used in this assay are obtained from the peritoneal cells of thioglycollate-induced (stimulated) and BCG-immunized (activated) mice; monocytes derived from human peripheral blood leukocytes (PBL), and the U937 macrophage-like cell line. The use of hapten-modified target cells facilitates a standardization of the ADCC assay and enables one to compare the cytotoxic activities of an effector cell population against different target cells modified with the same hapten and coated with the same antiserum. Cultured murine and human cell lines can be used as tumor targets. The human HSB cell line is most suitable as a target for both murine and human effector cells. New sera should be titered in the ADCC assay and the optimal dilution determined. It should be mentioned that mouse anti-TNP serum can be used effectively with human and murine effector cells, though other anti-TNP sera raised in other species may be more suitable with human effector cell. Peritoneal cells are obtained by the lavage of the peritoneal cavity with 10 ml EMEM containing 10 U heparin/ml.