Effects of TNF-α on murine T-cell IFN-γ secretion mediated by TNF-αRs. Recombinant TNF-α (50 ng/mL) were added to LN cells (LN) from WT B6 mice or TNFrsf1a^−/−1b^−/− mice under resting condition or activated by phorbol 12-myristate-13-acetate (PMA; 5 ng/mL) plus ionomycin (1 μM) overnight. Intracellular IFN-γ levels in CD4⁺ T cells (A) and CD8⁺ T cells (B) were examined by flow cytometry. Representative flow cytometry plots from 3 separate experiments are shown. (A-B) Top panels, Under resting condition; bottom panels, after stimulation with PMA plus ionomycin. (C) Transcriptome changes related to the impact of TNF-α on activated T cells. CD8⁺ T cells from B6 or TNFrsf1a^−/−1b^−/− mice were stimulated with PMA plus ionomycin in the presence of TNF-α (50 ng/mL) for 4 hours, and cells were then subjected to RNA extraction and complementary DNA (cDNA) synthesis. A PCR array focused on genes related to T-cell activation and anergy was performed. Genes with at least twofold differences between CD8⁺ T cells from B6 and TNFrsf1a^−/−1b^−/− mice are shown as a heatmap. Arrays were replicated from 2 different pools of CD8⁺ T cells. Red indicates high expression; blue, low expression. *P < .05.