Kinetics of cellular cytotoxicity mediated by a cloned human natural killer cell line.

ABSTRACT

The kinetics of natural killer (NK) cytotoxicity mediated by the cloned interleukin 2 (IL 2)-dependent human natural killer cell line NK3.3 has been investigated. This cloned cell line exhibits strong cytotoxic activity that is restricted to NK target cells. The initial rate of lysis of K-562 target cells by these cloned NK cells was, as anticipated, substantially greater than that previously reported for human peripheral blood NK cell preparations. However, in contrast to the kinetics of NK cytolysis mediated by fresh peripheral NK cells, the rate of 51Cr-release declined substantially within 1 to 3 h after initiation of assays involving NK3.3 cells and reached a plateau value in experiments conducted for longer periods. The data obtained suggest that NK3.3 cells cannot readily recycle and kill multiple target cells. Based on a model involving one lethal hit per active NK3.3 cell, estimates for the frequency of cytolytic cells in NK3.3 cultures were computed and compared to estimates obtained by the application of a kinetic model previously described. The cytotoxic activity of the NK3.3 cells was also found to be highly dependent on the conditions used for propagation and assay of these cells and, when cultured under "standard" conditions, only a fraction of the cloned NK3.3 cells was capable of effecting lysis of K-562 target cells. However, for the most optimal conditions developed to date, each NK cell killed an average of 1 to 2 target cells before inactivation. Although no significant differences in viability or growth rate were observed for the three different conditions described, up to 300-fold differences in lytic activity were observed. The observed strong dependence of the lytic function of NK3.3 cells on culture conditions should prove valuable for further investigations of mechanisms governing the regulation and function of human NK cells.