Figure 2.

Imaging cytometry of serum from Gambian HCC patients with high levels of ULBP1, stained for lipid membrane (BODIPY) and ULBP1. Representative examples of ULBP1 negative and positive exosomes shown in (A). Serum was separated into exosome and protein fractions by size exclusion column filtration and imaging cytometry of exosome-rich (B) and exosome depleted (C) fractions was performed. Exosomes gated as small, BODIPY positive events, a proportion of which are ULBP1 positive. ULBP1 concentration of whole serum, exosome rich and exosome depleted fractions was assessed by ELISA as in Figure 1 (D, n = 16). Flow cytometry of cell-surface expression of NKG2D on peripheral blood NK cells (E) and CD8 T cells (F) in a subset of Gambian patients with HCC or HBV. Ratio of NKG2D MFIs on paired peripheral and intrahepatic NK cells stratified by serum ULBP1 (G, n = 7). Cartoon depicting resection of tumour and liver tissue for short-term culture before ULBP1 ELISA of the conditioned media, ULBP1 concentration by ELISA of conditioned media from paired liver and tumour tissue (n = 1, H). Mean and SEM of all groups shown. Levels of significance: *p ≤ 0.05; **p ≤ 0.005; ***p ≤ 0.001; ****p ≤ 0.0001. Mann Whitney-U test was used for comparisons of two unpaired groups.