Figure 1

TIL activated by co-culture with M38 melanoma upregulate interferon signalling pathways. MSCs and HLA-A*0201+ TIL (TIL-H1 and TIL-A42) were co-cultured with HLA-A*0201⁺ M38 melanoma (n=12) or M28 melanoma, which lacked HLA-A*0201 (n=12); control TIL and TIL gene expression and supernatant protein levels were analysed. (A) Experimental design: MSCs were seeded at the bottom of a six-well plate; the melanoma cell lines, either M38 or M28, were seeded in the six-well transwells and placed above the MSCs. The TILs were added to the same transwells containing melanoma cells. (B) Ingenuity pathway analysis (IPA) of activated differentially expressed TIL genes showed that the expression of genes in the interferon signalling pathway changed, including IFN-γ, IRF1, JAK1, and STAT1. (C) In M38 melanoma-activated TIL IFN-γ increased 473-fold at the protein level and 20.24-fold at the gene level compared with control TIL. (D) Protein analysis shows the IFN-γ concentration in the co-culture supernatants (pg ml⁻¹). The pink bars represent supernatants from MSCs co-cultured with TIL-H1 and M38 melanoma cell lines and the yellow bars represent MSCs co-cultured with TIL-A42 and M38 melanoma cell lines.