Both mutations have been made in 129 ES-cells but backcrossed to C57BL/6J. The Il-10 gene is located on chromosome 1, whereas the Il-10r1 gene is located on chromosome 9. The regions Olaparib concentration flanking the mutation will still be derived from the 129 genome 15. Whether the presence of an alternative IL-10R ligand is the cause of the

differences observed in this study remains speculative. A similar phenomenon has been described for the IL-7−/−and IL-7R−/− mice, due to the binding of TSLP to IL-7R 16. However, IL-10−/− and IL-10R−/− mice always react in the same direction when compared with wt mice. In conclusion, as similarities prevail, the phenotype of IL-10R−/− mice is similar to the phenotype of IL-10−/− mice. Furthermore, these data confirm that IL-10 limits DSS-induced colitis. An induction

of IL-10 upon DSS exposure has been shown earlier 17. Monocytes/macrophages and/or neutrophils have been shown to be U0126 in vitro the main source for IL-10 in LPS-induced endotoxemia 2. We sought the main target cell of IL-10 in this model by analysing the different conditional IL-10R1 knock-out mice. Increased sensitivity to LPS was seen in IL-10−/−, IL-10R−/− and IL-10RFl/FllysM-Cre+ mice: An increase in the proinflammatory cytokines TNF-α, IL-1β and IL-12 was observed in IL-10−/− and IL-10R−/− compared with wt mice and IL-10RFl/FllysM-Cre+ compared with Cre-littermates. For IFN-γ, differences were significant between IL-10−/−/IL-10R−/− and wt and IL-10RFl/FllysM-Cre+ and wt mice respectively. IL-10RFl/FlCd4-Cre and IL-10RFl/FlCd19-Cre mice did not exhibit differences between Cre negative and positive littermates. Phosphoprotein phosphatase Results for IFN-γ and TNF-α are shown in Fig. 2B. IL-17 was expressed in the same pattern as TNF-α. Expression of IL-6 was highly induced by LPS in all

mouse strains. A slight increase was observed in IL-10−/− and IL-10R−/− compared with wt mice (Fig. 2C). A summary of all cytokines measured is shown in Supporting Information Table 1. IL-10 is crucial for the regulation of TLR-mediated innate immune responses. It inhibits the response to the TLR9 agonist CpG as well as to locally and systemically administered LPS 6, 18, 19. In particular, IL-10 produced by monocytes/macrophages and/or neutrophils was shown to be crucial for the regulation of the LPS-induced response, while T-cell-derived IL-10 is not necessary in this experimental setting 2, 20. In this regard, the observation that monocytes/macrophages and/or neutrophils are also the most important target cells of IL-10 in this model is not surprising. The presence of a self-regulatory loop in monocytes/macrophages or neutrophils, or the regulation of neutrophils by IL-10 produced by monocytes/macrophages or vice versa are probable models for the regulation of the systemic innate immune response to LPS. An autocrine loop in macrophages downregulating their own production of proinflammatory cytokines has been shown previously 21.