Additional file 1: of Cocultures of human colorectal tumor spheroids with immune cells reveal the therapeutic potential of MICA/B and NKG2A targeting for cancer treatment

Figure S1. Tregs infiltration in HT29 spheroids. Percentages of Foxp3+CD25+ Tregs among CD4+ T cells analyzed in Fig. 1c and d. Figure S2. IFNg blockade decreases spheroid infiltration and destruction by immune cells. (A) Pictures and analyses of (B) spheroid volume, (C) tumor cell apoptosis, and (D) spheroid infiltration 48h after coculturing HT29 spheroids with CD19-CD14- PBMCs in the presence or not of anti-IFNg blocking antibodies. Figure S3. T cell subsets and NKG2D expression by CD8 T cells after MICA/B treatment. (A) CD4 and (B) CD8 T cells proportions as well as (C) NKG2D expression by CD8 T cells relative to experiments in Fig. 5f to k. Figure S4. T and NK cells proportions and CD137 expression by CD8 T cells after combination therapy. Proportions of (A) T and NK cells, of (B) CD4 and CD8 T cells subsets and (C) CD137 expression by CD8 T cells relative to Fig. 6g to h. Figure S5. Pictures of primary CRC tumors cultures. Pictures of (A) primary CRC tumor cultured in adherent culture flasks and (B) tumor-derived spheroids used in autologous cocultures. Figure S6. Primary CRC-derived spheroids contains significant amount of EpCAM+ tumor cells. (A) Picture of primary CRC-derived spheroids and (B) flow cytometry or (C) IF analyses of EpCAM+ staining in the spheroids. Table S1. Clinical characteristics of the patients used for autologous cocultures. Table 2. Tumor cells content of the spheroids and T and NK cells composition of the TILs used for autologous cocultures. Percentages of tumor cells (EpCAM+CD45-) in patients-derived spheroids and percentages of NK cells (CD3e-CD56+) and T cells (overall CD3+, CD4 T cells CD3+CD4+CD8-, CD8 T cells CD3+CD4-CD8+) in respective autologous TILs used for cocultures. (DOCX 24846 kb)