2a–c). The nature and direction of the systemic immune response influences the pattern and severity of glomerular disease, therefore we measured immune responses directed against the nephritogenic antigen (i.e. sheep globulin). On day 21 systemic Th1 and Th17 cellular immune responses, assessed by antigen-stimulated splenocyte

cytokine production, were increased in STAT6–/– mice. Production of the key Th1-produced cytokine, IFN-γ, and key Th17-produced cytokine, IL-17A, were increased in STAT6–/– mice (Fig. 3a and b). In contrast, when assessing Th2 responses, there was no difference in IL-4 production (Fig. 3c); however, production of LBH589 datasheet IL-5 was decreased significantly in STAT6–/– mice (Fig. 3d). In addition, we measured IL-10 production from splenocyte cultures; however, levels were below the detection level of the assay in WT and STAT6–/– mice. These results demonstrated heightened Th1 and Th17 systemic immunity with a partial attenuation in Th2 responses. Humoral immune responses were assessed by measuring circulating antibody levels against the nephritogenic RXDX-106 mouse antigen. While WT mice with GN developed easily detectable antigen-specific humoral immune responses, there was a trend towards a decrease in measurable immunoglobulin (IgG) levels directed

against the nephritogenic antigen in STAT6–/– mice (Fig. 4a). Assessing IgG subtype production demonstrated a statistically significant decrease in antigen-specific IgG1 in STAT6–/– mice at serial dilutions, while production of antigen-specific IgG2b and IgG2c was unchanged. While the key Th1 (T-bet) [7] and Th17 (Roryt) [8] transcription factors influence the severity of renal injury in experimental crescentic GN, expression of these transcription factors peaks early in the disease process [7]. We measured expression of the key transcription

factors and cytokines after 6 days. No difference in splenic GATA3 expression was observed between WT and STAT6–/– mice. However, there was a significant increase in T-bet and Rorγ expression in STAT6–/– mice compared to WT mice given sheep anti-mouse GBM serum (Fig. 5a–c). There was no difference in splenocyte numbers in WT and STAT6–/– mice injected with sheep anti-mouse GBM serum (Fig. 6a). Antigen-stimulated cytokine production Dichloromethane dehalogenase demonstrated a trend towards increased production of IFN-γ and IL-17A in STAT6–/– mice (Fig. 6b and c). While production of IL-4 was detected readily in all samples, no difference was observed between WT and STAT6–/– mice (Fig. 6d). However, IL-5 production was decreased significantly in STAT6–/– mice on day 6 (Fig. 6e). There was no difference in antibody levels between WT and STAT6–/– mice on day 6; levels were not elevated compared to untreated mice. We analysed renal injury in WT and STAT6–/– mice 6 days after the administration of sheep anti-mouse GBM globulin.