This unit describes the monitoring and quantification of cellular cytotoxicity using a non-radioactive and real-time cytotoxic assay. The extent of target-cell lysis is monitored over time by imaging and quantifying live fluorescent target cells using a cell-imaging multimode reader. This assay is performed in a 96 well plate in optimized culture conditions at 37°C in the presence of 5% CO2 . The basic protocol describes natural killer cell-mediated cytotoxic assay that can be adapted to include antibodies blocking inhibitory NK receptors or triggering antibody-dependent cell-mediated cytotoxicity (ADCC). The assay is also suitable for antigen specific T-cell cytotoxic assays. Until now, the standard chromium 51 (51 Cr)-release assay has remained the sole sensitive assay but its major drawbacks include cost and hazard of handling radioactivity. The real-time cytotoxic assay is therefore an effective alternative providing a robust and sensitive assay that accurately monitors lysis of target cells over time. © 2017 by John Wiley & Sons, Inc.
Keywords: antibody-dependent cell-mediated cytotoxicity; cell cytotoxic assay; cytotoxic T cells; natural killer cells.
Copyright © 2017 John Wiley and Sons, Inc.