Fig. 2.

Validation of CD16 downregulation on NK cells within bladder tumours. CD16 marker expression on NK cells from human bladder tumours and matched blood was analysed by multiparametric flow cytometry. (a) Representative flow cytometry plots showing CD49a (ILC1-like marker) and CD49e (conventional NK cell marker) expression from one patient resected tumour and blood. (b) CD49a expression as a bar plot and (c) representative histogram plot for both tumour and blood NK (CD45⁺lin⁻CD56⁺NKp46⁺) cells. (d) CD16 expression as a bar plot and (e) representative histogram plot for both tumour and blood NK (CD45⁺lin⁻CD56⁺NKp46⁺) cell. (f) CD16 expression as a percentage of each gated cell type; conventional blood NK cell (CD45⁺lin⁻CD56⁺NKp46⁺CD49e⁺CD49a⁻) (cNK), tumour conventional NK cell (CD45⁺lin⁻CD56⁺NKp46⁺CD49e⁺CD49a⁻) and tumour ILC1-like NK cell (CD45⁺lin⁻CD56⁺NKp46⁺CD49e⁻CD49a⁺) displayed as bar plots and (g) representative contour plots. Data from 4 independent experiments (n = 4) and each symbol represents an individual patient bar graphs show mean value ± SEM Statistical P values determined by Mann–Whitney t test, where ∗P < 0.05. (H) Tumour bearing NSG IL-7/IL-15 KI mice were injected with isolated human PBMC and after 3 days blood and tumours were taken to analyse NK cell populations. Expression of CD16 on NK cells across each compartment. Data are from two separate pooled experiments, denoted by symbols (n = 9). Each symbol represents an individual mouse, bar graphs show mean value ± SEM. Statistical P values determined by Mann–Whitney t test, where ∗P < 0.05, and ∗∗P < 0.01.