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. 2019 Aug 14;9(20):5976–6001. doi: 10.7150/thno.33959

Figure 7.

Figure 7

Gal-3 is required for TLR-2-dependent activation of IDO1/KYN pathway in renal DCs and consequent generation of immunosuppressive Tregs. There was strong positive correlation between percentages of Gal-3+CD11c+DCs and TLR-2+CD11c+DCs in the kidneys (A). Representative flow cytometry dot plots showing expression of Gal-3 in TLR-2+ cells previously gated in the population of F4/80-CD11c+ renal DCs (B). Significantly lower number of TLR-2-expressing CD11c+DCs was observed in the kidneys of CDDP-treated Gal-3-/- mice compared to CDDP-treated WT animals, 72 h after CDDP injection (16 mg/kg body weight). There was strong positive correlation between percentages of IDO1+CD11c+DCs and TLR-2+CD11c+DCs in the kidneys (D). Representative flow cytometry dot plots showing expression of IDO1 in TLR-2+ cells previously gated in the population of F4/80-CD11c+ renal DCs (E). Representative dot plots showing that majority of TLR-2+Gal-3+IDO1+DCs express IL-10 (F). Significantly lower concentrations of KYN were noticed in supernatants of Gal-3-/-DCsPam3CSK4 and WTDCsPAM3CSK4+Davanat compared to WTDCsPam3CSK4 (G). Real-time PCR gene analysis (H), ELISA (I) and flow cytometry (J) results showing expression and production of inflammatory IFN-γ, IL-17, and anti-inflammatory IL-10 in Tregs (H, J) and appropriate supernatants (I) before and after culturing with WTDCsPam3CSK4, Gal-3-/-DCsPam3CSK4 or WT DCsPAM3CSK4+Davanat in contact-independent manner within transwell system (ratio between DCs and Tregs was 1:10). Real-time PCR gene analysis (K) and ELISA (L) results showing expression and production of inflammatory IFN-γ, IL-17 and anti-inflammatory IL-10 in LPS-activated neutrophils that were cultured with Tregs previously stimulated by WTDCsPam3CSK4, Gal-3-/-DCsPam3CSK4 or WT DCsPAM3CSK4+Davanat. Transwell systems were used to separate neutrophils and Tregs and ratio between neutrophils and Tregs was 10:1. Data from two individual experiments with 8 mice per group are shown as Mean ± SEM; *p<0.05, **p<0.01;***p<0.001.