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. 2018 Feb 13;68(2):335–346. doi: 10.1136/gutjnl-2017-315485

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Immune profiles of tumour infiltrating leucocytes (TILs) isolated from Yttrium-90 (Y90)-radioembolisation (RE)-treated and treatment-naïve tumours. (A) Samples collection and analysis pipeline. Peripheral blood mononuclear cells (PBMCs) were collected before (pre-Y90) and at various time points after Y90-RE (post-Y90) (n=31 patients). TILs were collected from resected hepatocellular carcinoma (HCC) tumours from post-Y90-RE (downstaged on therapy) or treatment-naïve patients, control (Ctl) (n=7 for each group). time-of-flight mass-cytometry (CyTOF) was used to analyse both the PBMCs and TILs and next-generation sequencing (NGS) was performed on tumour tissues from post-Y90-RE and treatment-naïve patients (n=4 for each group). (B) Two-dimensional (2D) heat map showing the differential expression of immune markers by nodes enriched in TILs isolated from post-Y90-RE (red bar) or treatment-naïve (Ctl; green bar) HCC tumours. Enriched immune subsets in TILs from post-Y90-RE were CD56⁺natural killer (NK) cells, CD8⁺CD56⁺ NKT cells, CD8⁺ T and CD4⁺ T cells while regulatory T, Treg cells were enriched in TILs from Ctl HCC (colour-coded lines). n=7 each group. (C) 2D representation of granzyme B (GB) and Tim-3 expression on TILs isolated from post-Y90-RE (Y90) and Ctl HCC tumorstumours. Images were generated using MARVis software. () Representative plots showing the gating of GB on CD8⁺ T cells from post-Y90-RE (Y90) or Ctl TILs (left panel). Percentage of GB⁺CD8⁺ and Tim-3⁺CD8⁺ T cells from post-Y90-RE and Ctl TILs (right panel). () Percentage of CD56⁺ NK cells, CD8⁺CD56⁺ NKT cells, GB⁺CD56⁺ NK cells and GB⁺CD8⁺CD56⁺ NKT cells from post-Y90-RE (Y90) and Ctl TILs. Graphical data represent the means±SD and were analysed by unpaired Student’s t-test. *P<0.05 and **P<0.01.