Real-time profiling of NK cell killing of human astrocytes using xCELLigence technology.

We have conducted the first profiling of human Natural Killer (NK) cell mediated killing of astrocytes using xCELLigence technology. The sensitivity and applicability of xCELLigence was compared to lactate dehydrogenase (LDH) release and time-lapsed microscopy to validate the killing events. The xCELLigence technology uses electrical impedance measurements from adherent cells and converts into Cell Index (CI). NK cells did not register any Cell Index signal directly, therefore all changes in Cell Index are a direct measure of astrocyte responses. Astrocytes are insensitive to basal NK cells (non-activated NKs). Whereas NK cells activated by IL-2 prior to culture with targets rapidly kill astrocytes. This observation was supported by all methods of analysis. Using the xCELLigence we were able to monitor the longer term killing profile. This demonstrated that at all NK ratios, death was achieved if given long enough. In addition, the development of the killing phenotype was investigated by inducing lymphokine activated killing with IL-2 in the presence of the target astrocytes. In this paradigm of killing, the xCELLigence was the only assay suitable due to the prolonged time-course required for killing, which required 4-5 days to achieve maximal killing (100%). This suggested that the astrocytes can directly suppress the killing activity of the NK cells. These data highlight the sensitivity, applicability and profiling power of the xCELLigence system and support its use for further investigation of NK-killing of healthy and/or tumourogenic astrocytic cells.