Figure 7. Adipose iNKT cells enhance the suppressive ability of T_regs and have similar functions to T_regs.

(a) Dot plots and graph of Klrg1 surface expression on splenic T_reg cells (CD3⁺CD4⁺Foxp3⁺) in spleen 3 days after PBS or αGalCer treatment, or in WT adipose tissue 3 days after PBS or αGalCer treatment, or in adipose tissue of CD1d^−/− mice (n=5 per group). (b) Intracellular IL-10 production in vivo by T_reg cells in spleen (first 2 bars) and adipose tissue (last 2 bars) after treatment in vivo with αGalCer (n=4). (c) Basal intracellular IL-10 production in vivo by adipose T_regs in WT and Ja18^−/− mice (n=6). (d) 10,000 T_regs were isolated from adipose tissue that had been treated with PBS or αGalCer for 3 days (pooled from 5 mice per treatment group), and cultured with peritoneal macrophages. MFI of CD301 and CD206 levels were measured on cultured macrophages (n=3 experiments). (e) Klrg1, ICOS and PD-1 surface expression on iNKT cells from spleen and adipose tissue (n=5–6). (f) Peritoneal macrophages were isolated from WT mice and cultured for 48hrs with either iNKT cells or T_reg cells from adipose tissue or media alone. After 48hrs of culture, macrophages were stained for flow cytometric analysis of CD206/MMR and CD301 levels (n=2 independent experiments). (g) Dot plots and graphs of intracellular iNOS levels in peritoneal macrophages from obese mice cultured with adipose T_reg cells and iNKT cells for 24 hours. Statistical comparisons using t-tests or ANOVA for group of 3 with Tukey post-hoc test. *p<0.05, **p=<0.01, ***p<0.001.