A strategy for evaluating lymphokine activation and novel monoclonal antibodies in antibody-dependent cell-mediated cytotoxicity and effector cell retargeting assays.

Abstract

As novel antibody therapeutics are developed for different malignancies and require evaluation with cells previously uncharacterized as antibody-dependent cell-mediated cytotoxicity (ADCC) targets, efficient description of key parameters of the assay system expedites the preclinical assessment. A strategy is presented to define the behavior of cell lines or cell cultures as targets in ADCC assays, with emphasis on cytokine activation of effectors and attention to contributions of natural killer cells. Features of the target cell, the effector cell, and the assay itself are separately assessed. Target cells are evaluated for the kinetics of chromium labelling and release, and positive and negative control antibodies are selected. Effector cells are evaluated in ADCC for the impact of different donor sources, storage conditions, lymphokine concentration and duration of activation. The assay itself is assessed for the impact of the type of liquid medium, incubation duration, and effector-to-target ratio. Representative data are presented with a model human malignant T cell line, HuT102.