5A and data not shown). OVA-specific Th2-cell dependent IgG1 and IgE were detected in the serum of mice upon alum/OVA sensitization and antigen challenges. Surprisingly, no change was detectable Alisertib research buy at the OVA-specific Ig levels when mice were pretreated with the differentially matured and OVA-loaded DCs (Fig. 5B). Together, MyD88-dependent T. brucei-derived VSG antigens or nonTLR-dependent TNF conditioning of DCs did not alter subsequent Th2-cell driven allergic asthma. EAE serves as a common murine model for the early phases of multiple sclerosis, which can be achieved by immunizing

mice with the auto-antigen MOG in CFA. Mice develop MOG-reactive pathogenic Th1 and Th17 cells, which then infiltrate into the MK-2206 in vivo CNS and cause inflammatory edema leading to the reversible paralysis symptoms 43. Previously, we have shown that repeated injections of DCs stimulated with TNF and loaded with MOG-peptide suppressed EAE, partially by creating a Th2/Tr1 cytokine environment including immune deviation and IL-10-mediated suppression 23, 33. We therefore wanted to analyze how the partial DC maturation stages induced by TLR-dependent or independent

stimuli would modulate the autoimmune disease EAE. To detect whether the DC injections ameliorate or worsen the disease, we switched the amounts per DC injection from 3 to 3.5×106 cells, which is the fully protective protocol 23, 33, 44 to 2–2.5×106 cells, which leads to about 50% reduced clinical score 44. Three i.v. injections of suboptimal amounts of MOG-loaded TNF-matured DCs protected mice partially from EAE as 10 out of 15 mice Methocarbamol developed clinical symptoms and mice only reached a mean maximum score of 1.850±0.944 (Fig. 6A and B). Surprisingly, mice which received three injections of DCs matured with the T. brucei antigens mfVSG or MiTat1.5 sVSG were also partially protected from EAE as 8 out of 12 and 13 out of 19 mice developed signs of EAE, respectively (Fig. 6A and B). Together, our data indicate that all partially mature DCs protected mice

to a similar extent from EAE. As published previously 33, protection from EAE by TNF-matured DCs required activation of IL-10+ IL-13+ cytokine-producing CD4+ Th2/Tr1 cells. IL-4 is also produced but immediately consumed in normal mice and only detectable in IL-4R-deficient mice 33. We therefore assessed how the differentially matured DCs influenced the T-cell cytokine profile of the spleens as detected after MOG peptide restimulation and cytokine analysis. The cytokine profile of T cells from untreated mice typically consists of high amounts of proinflammatory IFN-γ and IL-17 but low amounts of IL-10 and IL-13. In contrast, this pattern becomes inverted in mice, which received repetitive injections of TNF-matured DCs 23, 33.