Fig. 1: Melanoma fractions resistant to antigen-specific T cells.
From: Reversal of pre-existing NGFR-driven tumor and immune therapy resistance
a Graphic overview of the generation of T cell-resistant (TR) cell lines. b Colony formation assay of three parental cell lines (par) and their TR counterparts, treated with either control (Ctrl) or MART-1 T cells for 24 h in a 1:1 ratio and recovered for 3 days. Quantification in Supplementary Fig. 1a. c Tumor growth of melanoma cell lines D10 and the T cell-resistant counterpart, D10-TR. Randomization occurred on day 10 (indicated by dotted line). n = 7 per group except n = 6 for D10 + MART-1 T cells. Statistical analysis by Kruskal–Wallis test on day 21; *p < 0.05. d mRNA expression levels of MLANA in parental versus TR cell lines. Pooled data of six independent cell line pairs, lines indicate each paired parental and TR cell line. One experiment of three pooled technical replicates is shown; the data are reproduced in two independent replicates (available in Source data). e Cell viability after T cell attack of M009R.X1.CL cells for CDKR24C and NY-ESO-1 TCRs. An experiment of two independent replicates with three technical replicates is shown (other replicate can be found in Source data). Statistical analysis by unpaired t-test; **p < 0.01. f Gene expression changes and average signature scores in TR cells vs. parental cells, normalized per cell line. Gene lists for different subtypes/cell states were derived from Tsoi et al. (see Methods section). g Flow cytometry quantification of AXL and NGFR expression in six matched cell lines. Error bars indicate S.D. of four independent replicates. Statistical analysis by Mann–Whitney test; *p < 0.05, ns not significant. h Immunohistochemistry analysis of NGFR on human melanoma, D10 tumors and D10-TR tumors. Scale bars indicate 50 μm. i FACSort lay-out of NGFRlow and NGFRhi cells. j Quantification of cell viability after MART-1 T cell attack in three cell lines, similarly sorted as in i. Data of two independent replicates with two and three technical replicates, respectively. Statistical analysis by Mann–Whitney test; *p < 0.05, **p < 0.01. All error bars in this figure represent S.D. Source data are provided as a Source Data file.
