51Cr-release to monitor NK cell cytotoxicity

Natural killer (NK) cells are cytotoxic lymphocytes that belong to the innate lymphoid cells. They have the ability to attack tumor cells that lack ligands of inhibitory NK receptors and/or express ligands of activating NK receptors. The most important ligands of inhibitory NK receptors are major histocompatibility complex (MHC) class I molecules, which are down regulated on many tumor cells. Several ligands of activating NK receptors, in contrast, are up regulated on tumor cells due to cellular and genotoxic stress. Due to these features, NK cells constitute an important element of the tumor immunosurveillance and are explored as a cellular tool for tumor immunotherapy. Therefore, it is important to be able to monitor the cytotoxic activity of NK cells reliably in preclinical animal models as well as in patients. It is also of interest to determine the susceptibility of tumor cells toward NK cell-mediated cellular cytotoxicity. The Cr-51-release assay monitors the lysis of target cells by NK cells and is suitable to address both questions. We describe here the experimental set-up of this classic cytotoxicity assay in detail.