A Real‐Time Cytotoxicity Assay as an Alternative to the Standard Chromium‐51 Release Assay for Measurement of Human NK and T Cell Cytotoxic Activity. (13th February 2018)

• Record Type: Journal Article
• Authors: Fassy, Julien; Tsalkitzi, Kyriaki; Goncalves‐Maia, Maria; Braud, Véronique M.
• Editors: Coligan, John E.; Bierer, Barbara E.; Margulies, David H.; Shevach, Ethan M.; Strober, Warren
• Is Part Of: Current protocols in immunology. Volume 118(2017)
• Page Start: 7.42.1
• Page End: 7.42.12
• Publication Date: 2018-02-13
• Subjects: cell cytotoxic assay -- antibody‐dependent cell‐mediated cytotoxicity -- natural killer cells -- cytotoxic T cells; Immunology -- Laboratory manuals; Medical protocols; Immunologic Techniques; Medical protocols; Immunology; Laboratory manuals; Reference works; 571.96028
• DOI: 10.1002/cpim.28
• Languages: English
• ISSNs: 1934-3671

Abstract

Abstract: 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.