Apc tcrvβ11

Invariant NKT cells were stained using PE-TCRvα24 (Beckman Coulter, Marseille, France) and APC-TCRvβ11 (BD Biosciences) mAbs. IgG1 mAbs (Beckman Coulter or BD Biosciences) were used as isotype controls. To detect intracellular cytokines, cells were fixed and permeabilized using Cytofix/Cytoperm kits (eBiosciences) according to the manufacturer's instructions. These cells were incubated with FITC-IL-4 (BD Biosciences), FITC-IFN-γ (BD Biosciences), FITC-IL-17 (Biolegend), or FITC-IL-10 mAbs (BD Biosciences). IgG1 isotype controls (Biolegend or BD Biosciences) were used. After gating on lymphocytes and Vα24⁺Vβ11⁺ iNKT cells, IL-4⁺, IFN-γ⁺, IL-17⁺, or IL-10⁺ cells were expressed as a percentage of Vα24⁺Vβ11⁺ iNKT cells (Fig. 2A).

To quantify Foxp3⁺ iNKT cells, PBMCs were permeabilized using a Foxp3 staining buffer kit (eBioscience) and stained with FITC-TCRvα24 (Beckman Coulter), APC-TCRvβ11 (BD Biosciences), and PE-Foxp3 (eBioscience) mAbs. IgG2aκ (eBioscience) was used as an isotype control for Foxp3 mAb. After gating on lymphocytes and Vα24⁺Vβ11⁺ iNKT cells, Foxp3⁺ cells were expressed as a percentage of Vα24⁺Vβ11⁺ iNKT cells.