Anti pd 1 clone j43

C57/BL6 mice were treated with 200 µg anti–PD-1 (clone J43; Bio X-Cell), 50 µg anti–4-1BB (clone 3H3; Bio X-Cell), or respective hamster or rat isotype controls, intraperitoneally every other day for 3 d (in non–tumor-bearing experiments) or every other day for the duration of the experiment (in tumor experiments). For tumor growth experiments, mice were injected intradermally with 100,000 B16-F10 melanoma cells. When tumors were between 1 and 3 mm in any direction (typically day 6–7), mice began receiving immunotherapy. For TIL analysis experiments, tumors were allowed to reach 5–7 mm in any direction before initiation of immunotherapy in order to obtain sufficient cell numbers. For adoptive cell therapy experiments, mice received B16^OVA intradermally. OT-I T cells were activated in vitro by using either SIINFEKL peptide (500 nM) or anti-CD3 stimulation in combination with anti-CD28 or anti-–4-1BB, followed by an expansion in 50 U/ml recombinant mouse IL-2. When tumors were between 1 and 3 mm in any direction (typically day 7–8), mice received an intravenous adoptive transfer of 2 × 10⁶ or 5 × 10⁶ OT-I cells. In systemic immunotherapy experiments, mice then began receiving 50 µg anti–4-1BB or its isotype control intraperitoneally.