Fig. 2: Decitabine enhances γδ T cell-mediated cytolysis of lung cancer cells.

a Bar graphs showing percentages of γδ T cell subsets in the CD3+ population from the peripheral blood of a healthy donor following ex vivo expansion. Data are presented as mean ± standard error of the mean (SEM). n = 3, independent experiments. b Overlay of tSNE maps of CD3+ T cells from PBMC at baseline and after ex vivo expansion analyzed by mass cytometry. Each dot represents a single cell. The color represents the expression level of the indicated markers. Red is high, and blue is low. c Pearson correlation of markers expressed in ex vivo expanded γδ T cells analyzed by mass cytometry. d Representative flow cytometric analysis of annexin V and propidium iodide (PI) apoptosis assays in human lung cancer cell lines upon treatments with 100 nM decitabine (DAC) alone, γδ T cells alone or DAC/γδ T cell combination. The effector to target (E:T) ratio is 3:1. Gating strategies are shown in Supplementary Fig. 16. e Dot plots showing three biological replicates (mean ± SEM) of apoptosis assays described in d. f Annexin V and propidium iodide apoptosis assays of a patient-derived lung cancer cell line from malignant pleural effusion, PD#0899 (mean ± SEM, n = 3, independent experiments). g Transwell migration assays of γδ T cells (upper chamber) towards mock- or DAC-treated lung cancer cells (lower chamber). Numbers of γδ T cells in the lower chambers are counted per high power field. Representative images of γδ T cells (Hoechst 33342-labeled; green) and lung cancer cells (Calcein-retained; red) in the lower chambers are shown. Data are presented as mean ± standard deviation (SD). n = 15 high power fields, over three independent experiments. Scale bar: 100 μm. The p value is calculated by one-way ANOVA with Tukey’s multiple comparison test (panels a, e and f) or the two-sided Mann–Whitney test (g).