05), IFNγ- (P < 0.01), and IL-17- (P < 0.05) and -4-producing CD4+ cells (P < 0.01) in Gal-3−/−, when compared buy Y-27632 to WT, mice 8 hours after Con A injection (Fig. 3). Furthermore, the total number of IL-12-producing CD11c+ DCs as well as IFNγ- and IL-4-producing NKT cells were significantly lower (P < 0.05) in livers of Gal-3−/−, when compared to WT, mice (Fig. 4). There was no significant difference in the total number of IFNγ-producing CD8+ T and NK cells and IL-10-producing CD11c+ DCs (data

not shown) between WT and Gal-3−/− mice. Interestingly, the total number of IL-10-producing CD4+ T cells and F4/80+ macrophages was significantly higher (P < 0.05) in livers of Gal-3−/−, compared to WT, mice (Figs. 3 and 5). Additionally, the ratio between the total number of IL-10- and IFNγ-producing CD4+ T cells was significantly higher (P < 0.05) in livers of Con A–treated Gal-3−/−, compared to WT, mice (4.53 ± 0.74 Gal3−/− versus 2.35 ± 0.56 WT). We did not find any difference in the total number of liver F4/80+ macrophages between WT and Gal-3−/−

mice, but we noticed a significant difference in the total number of IL-10-producing F4/80+ cells (Fig. 5A). We found a significantly higher percentage and total number of F4/80+ CD206+ alternatively activated (i.e., M2-polarized) macrophages in livers of Gal-3−/−, compared to WT, mice (Fig. 5A). Thus, it appears that Gal-3 deletion favors the differentiation of IL-10-producing macrophages. Talazoparib nmr We assumed that apoptosis of infiltrating cells may contribute to the lower number of MNCs in livers of Gal-3−/− mice. Indeed, we found enhanced apoptosis of liver-infiltrating MNCs and splenocytes in Gal-3−/−,

compared to WT, mice (Fig. 5B; Supporting Fig. 5) 8 hours after Con A injection. Both in livers and spleens, the majority of MNCs were in the stage MCE of late apoptosis (Annexin V+ propidium iodide [PI]+ cells; Fig. 5B; Supporting Fig. 5). Significantly higher percentages of Annexin V+ PI+ liver-infiltrating MNCs (P < 0.05) and splenocytes (P < 0.05) were observed in Gal-3−/−, mice compared to WT, mice (percentage of apoptotic cells in liver: 34% Gal3−/− versus 18.8% WT; in spleen: 38.7% Gal3−/− versus 17.3% WT). To further elucidate the role of Gal-3 in Con A–induced liver injury, we pretreated WT C57BL/6 mice with TD139, competing for the saccharide-binding site, 2 hours before and immediately after Con A injection. We found that the administration of TD139 prevented the increase of serum liver transaminases (Fig. 6A). This finding was consistent with scarce necrotic areas observed in the livers of pretreated animals, in contrast to significantly larger necrotic areas in liver parenchyma of mice treated with Con A and vehicle (Fig. 6B). IP injection of TD139 in Con A–untreated animals did not alter the serum level of liver enzymes (data not shown).