Anti ifn γ antibody

Anti-mouse CD152 (anti-CTLA-4) (Clone 9H10, GoInVivo^™, BioLegend, CA, USA) was administered i.p. at 5μg/g mouse body weight on days 8, 11, and 14 in 100μL PBS.(21 (link)) Anti-mouse CD279 (anti-PD-1) antibody (Clone RMP1-14, GoInVivo^™, BioLegend, CA, USA) was administered i.p. at 5μg/g mouse body weight on days 8, 11, and 14 in 100μL PBS. Anti-IFN-γ antibody (Clone XMG1.2, BioXCell, NH, USA) was administered i.p. at 25μg/g mouse in 200μL PBS. (22 (link)) Corresponding isotype controls were used: mouse IgG1 Isotype Ctrl Antibody and rat IgG2a Isotype Ctrl Antibody (BioLegend, GoInVivo^™). 1-methyl-D-tryptophan (Sigma-Aldrich, MO, USA) was administered ad lib in drinking water of mice as previously described.(14 (link)) Mice drank approximately 2.5–3.5mL of 1-D-MT supplemented water per day. Epacadostat (Selleckchem) was administered 300mg/kg.(23 (link))

For T_H0, T_H1 or induced T_reg cell differentiation, naïve CD4⁺ T cells were stimulated with 5 μg/ml each of plate-bound α-CD3/CD28 antibodies in the presence of human IL-2 (100 U/ml; PeproTech) for T_H0 polarizing; human IL-2 (100 U/ml) plus mouse IL-12 p40 (0.5 ng/ml; BD Biosciences) for T_H1 polarizing; or human IL-2 (100 U/ml) plus human TGF-β (0.5 ng/ml; PeproTech) for T_reg polarizing for 2.5 d, and rested in complete Click’s medium (IrvineScientific) supplemented with 10% (vol/vol) FBS and 1% (vol/vol) penicillin-streptomycin and the cytokine combinations indicated above for 3 d. For T_H2 or T_H17 cell differentiation, naïve CD4⁺ T cells were stimulated with 2 μg/ml each of soluble α-CD3/CD28 antibodies and irradiated antigen presenting cells (T cell-depleted splenocytes) in the presence of human IL-2 (100 U/ml), mouse IL-4 (10 ng/ml; R&D systems) and anti-IFN-γ antibody (10 μg/mL; Bio X Cell) for T_H2 polarizing; or human TGF-β (2 ng/ml; PeproTech) plus mouse IL-6 (20 ng/m; BD Biosciences) for T_H17 polarizing in complete Click’s medium for 5.5 d.