Fas ligand and lytic granule differentially control cytotoxic dynamics of natural killer cell against cancer target

Interaction dynamics between Natural Killer (NK) cells and cancer targets have been the topic of many previous investigations, but the underlying rate-limiting kinetics and heterogeneity remain poorly understood. In this study, using quantitative single cell microscopy assay, we elucidate the differential dynamic control of NK-cancer cell interaction by multiple cytotoxic pathways. We found primary human NK cell, unlike NK cell line, killed adherent cancer target mainly by lytic granule-independent mechanism, in particular through Fas ligand (FasL). And the distinct kinetics of FasL and lytic granule pathway resulted in significant cell-to-cell variability. Killing by FasL occurred slowly, requiring transient, often multiple NK-cancer cell conjugations that gradually activated caspase-8, while lytic granule triggered rapid cytotoxicity by a switch-like induction of granzyme-B upon a single, prolonged conjugation. Moreover, interleukin 2 was observed to enhance both cytotoxic mechanisms by promoting target recognition by NK cell and increasing NK-cancer cell interaction frequency. Our results not only identify the key points of variation in the rate-limiting kinetics of NK-cancer cell cytotoxic interaction but also point to the importance of non-lytic granule mechanism for developing NK cell therapy.