To further understand the mechanisms through which IL-4 contributes to α-Galcer-induced liver injury, we examined the role of STAT6 in this model. As illustrated in Fig. 4A, STAT6 was activated in neutrophils from the livers of α-Galcer-treated WT mice, whereas this activation was diminished in neutrophils from α-Galcer-treated IL-4−/− mice. This result suggests that IL-4 is responsible for the observed STAT6 activation in neutrophils. In agreement with

the data from IL-4−/− mice, STAT6−/− mice had lower serum levels of ALT and AST (Fig. 4B), fewer inflammatory CX-5461 manufacturer foci (Supporting Fig. 2), and a reduced number of neutrophils in the liver (Fig. 4C) compared to WT mice post-α-Galcer administration. In addition, neutrophils from α-Galcer-treated STAT6−/− mice demonstrated higher levels of apoptosis than those from WT mice (Fig. 4D). Collectively, our findings suggest that IL-4/STAT6 inhibit

neutrophil apoptosis. To understand the mechanisms underlying the IL-4/STAT6-mediated inhibition of neutrophil apoptosis, we investigated the expression of NVP-LDE225 antiapoptotic genes in these cells and identified that the expression of survivin and Bcl-2 was significantly up-regulated in hepatic neutrophils from α-Galcer-treated WT mice, whereas this up-regulation was reduced in hepatic neutrophils from α-Galcer-treated IL-4−/− or STAT6−/− mice (Fig. 4E). The finding that deletion of IL-4 abolished α-Galcer-induced hepatitis cannot explain the exacerbated α-Galcer-induced liver injury observed in IL-4−/−IFN-γ−/− dKO mice. To further understand the mechanisms by which IL-4−/−IFN-γ−/− dKO mice are more susceptible to α-Galcer-induced hepatitis, we examined this model in IFN-γ−/− or IFNGR−/− mice. As

illustrated in Fig. 5A, IFN-γ−/− or IFNGR−/− mice were more sensitive to α-Galcer-induced liver injury, as reflected by the higher levels of serum ALT and AST than WT mice. selleck chemical In agreement with the biochemical data, histological examination, as shown in Fig. 5B, confirmed more severe liver injury and inflammation (larger area of necrosis and a larger number of inflammatory foci) in IFN-γ−/− and IFNGR−/− mice at both 16 hours and 72 hours after α-Galcer administration than in WT mice. In addition, the number of MPO+ neutrophils was higher in the livers of IFN-γ−/− or IFNGR−/− mice post-α-Galcer injection (Fig. 5B). Because it has been shown that NKT and NK cells can kill hepatocytes and contribute to liver injury,[18, 19] we hypothesized that the differences in α-Galcer-induced liver injury in WT and IFN-γ−/− mice were due to varying degrees of NKT and NK activation. The data in Supporting Fig.