Fig. 3

Vγ2Vδ2 T cells expanded by pulse zoledronate stimulation exhibit similar cytotoxicity against pamidronate-treated PC-3 cancer cells compared with Vγ2Vδ2 T cells expanded by continuous stimulation. Vγ2Vδ2 T cells were expanded as described in Fig. 2. Vγ2Vδ2 T cells expanded by continuous or by pulse zoledronate stimulation were then purified for use. PC-3 cancer cells were treated overnight with pamidronate (200 μM) and then washed twice. To assess cytotoxicity, the FATAL assay was used. Pamidronate-treated PC-3 cells were stained with PHK-26 and CFSE and incubated with purified Vγ2Vδ2 T cells for 5 h at various E:T ratios in duplicate. Cells were then fixed with 1% paraformaldehyde followed by flow cytometric analysis. a CFSE levels of PC-3 prostate cancer cells incubated with varying numbers of purified Vγ2Vδ2 T cells expanded by continuous or pulse zoledronate stimulation. b Cytolytic activity of Vγ2Vδ2 T cells expanded either by continuous or pulse zoledronate stimulation against pamidronate-treated PC-3 cancer cells as determined by the FATAL assay. Representative of two experiments