T cell–derived cytokines drive immune evasion.

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T cell–derived cytokines drive immune evasion. T cell–derived cytokines drive immune evasion. (A) RNA-seq (triplicate samples) of MC38Ova cells that were cocultured with OT-I T cells for 6 hours. (B) Volcano plot of top regulated genes from (A). (C) GSEA from (A). (D) MC38Ova cells were treated with supernatant (6 hours) derived from an overnight MC38Ova coculture with OT-I T cells (1:1 E:T ratio) followed by 3′ RNA-seq. GSEA is displayed. (E) MC38Ova killing assay with Prf1−/− OT-I T cells ± α-TNF or α–IFN-γ (25). MC38Ova killing assay using supernatant from an MC38Ova/vec coculture with OT-I T cells ± α-TNF or α–IFN-γ (25 μg/ml). MC38 Ova killing assay upon TNF or IFN-γ treatment at the indicated concentrations for 18 hours. (F) Custom sgRNA pool screen in MC38Ova cells. Cells were treated with TNF (1 ng/ml) three consecutive times. (G) Comparison of the top scoring genes from (F) to the OT-I T cell screen described in Fig. 1C. (H and I) MC38 and B16 cells carrying the custom gRNA library were treated with IFN-γ overnight and then cell-sorted for MHC-I negatives (repeated three times). Cells were then sequenced for gRNA enrichment. (J) Selected gRNAs from the MC38 screen in (H). Conor J. Kearney et al. Sci. Immunol. 2018;3:eaar3451 Copyright © 2018 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works