Figure 7
MAIT and NK cells kill myeloma cells with equivalent kinetics. Purified MAIT cells were loaded with Fluo-4-AM and co-cultured 1:1 with RPMI-8226 myeloma cells in media supplemented with 100 nM 5-OP-RU and 100uM propidium iodide. To compare MAIT cell killing kinetics of target cells, comparator co-cultures included purified NK cells co-cultured with RPMI-8226 or K562 (E:T ratio 1:1). Time lapse live video data was acquired on a Leica SP5 confocal microscope and analysed using Metamorph Imaging series 7 software. Data is presented in: (A) Montage of a single MAIT killing event with timestamps for MAIT attachment, MAIT Ca2+ flux, and myeloma cell blebbing (apoptosis), or (B) As collated data of individual killing events and shows the time taken (minutes) from initial contact of effector cell to target cell blebbing (apoptosis) for NK cells with K562 cells (n = 44, left), NK cells with RPMI-8226 cells (n = 28, middle) or MAIT cells with RPMI-8226 (n = 48, right). Statistical analysis (student’s t-test) was performed between test and control groups, (ns) not significant. (C) A video montage shows an individual MAIT cell which undergoes attachment, Ca2+ flux, and sequential killing of two 5-OP-RU pulsed RPMI-8226 cells.
