We also detected a small, yet reproducible, population of IL-13+, IL-17A+, and IFN-γ+ “triple-positive” cells, thereby demonstrating that, in some inflammatory settings, IL-13 can be produced by unorthodox T-cell “subsets” (Fig. 1E and F). To confirm our flow cytometry studies, we purified donor T cells from immunized Balb/c and sOva Rag2−/− hosts, then measured cytokines and TFs by PCR. Consistent with our protein measurements, we found that www.selleckchem.com/products/sorafenib.html both groups expressed high levels of IL-13, IFN-γ, and IL-17A mRNA. T-bet and RORγT, the signature TFs for Th1 and Th17 cells, were similarly

abundant in both groups, but GATA-3 and IL-4 were much more abundant in the immunized group, thus highlighting the atypical nature of the IL-13 response in sOva Rag2−/− hosts. To further explore the role of TFs in IL-13-producing Th1, Th2, and Th17 cells, we turned to the DSS colitis model. First, we used flow cytometry to measure TF levels in CD4+ TCRβ+ IL-13+ T cells, finding that many coexpressed high levels of GATA-3, T-bet, or RORγT (Fig. 1F). Next, we did the converse experiment and measured IL-13 and TFs within IL-4+, IFN-γ+, or IL-17+ cells. As expected, we found that a large percentage of IL-4+ cells expressed high levels of IL-13 and GATA-3, thus representing “classical” Th2-type effectors. We could also detect IFN-γ+ Cytoskeletal Signaling inhibitor cells capable producing IL-13. These were largely

T-betlow, which suggests they could be in a transitional state, either coming from or moving toward Tbethigh Th1 effectors. A smaller population of IL-17A/IL-13 double-positive cells was observed but, in this case, they were RORγThigh, leading us to conclude that IL-13 can be produced by bonafide Th17-type effectors (Fig. 2B and Supporting Information Fig. 4). To ask whether canonical Th2-type signals are required for the development

of IL-13-producing Th1 and Th17 cells, we transferred IL-4Rα- or STAT6-deficient donor T cells into sOva Rag2−/− hosts. Consistent with the known ability of Th2-type cytokines to suppress Th1 and Th17 responses [2], we found that IL-4Rα−/− and STAT6−/− donors produced Cyclic nucleotide phosphodiesterase more IFN-γ and IL-17 than WT counterparts. More importantly, despite a slight reduction in total IL-13+ cells, IL-13-producing Th1 and Th17 cells, we still generated, using IL-4Rα−/− or STAT6−/−, donors, which demonstrates that, under conditions of acute inflammation, Th1 and Th17 cells can produce IL-13 in the absence of IL-4Rα or STAT6 (Fig. 2C). To investigate the function of IL-13 and Th2-type cytokines in the context of Th1- and Th17-mediated inflammation, we paired WT and IL-4Rα-deficient donors together with WT and IL-4Rα-deficient sOva Rag2−/− hosts, creating a system where either the T cells (donor) or non-T cells (host) were lacking IL-4Rα. As expected, we found that the pairing of WT donors and WT hosts resulted in lethal autoimmune disease between 7 and 10 days posttransfer.