Blockade of the Tim-3 and PD-1 signaling pathways restores IFN-γ production. TILs were harvested from CT26 tumor-bearing mice and cultured in vitro in the presence of soluble anti-CD3 and anti–Tim-3, anti–PD-L1, anti–Tim-3 plus anti–PD-L1, or control immunoglobulins. After 96 h, culture supernatant was collected and IFN-γ measured by cytometric bead array. Data are expressed as the difference in cytokine production over that observed in cultures with control immunoglobulins. Data shown are from three independent TILs samples from two independent experiments.