2C). These genes are largely overlapping with those reported by others before as a typical inflammatory pattern for DCs 40 and thereby indicate the reliability of our microarray approach. The different intensities of induction between TNF/mfVSG/MiTat1.5 sVSG and LPS further strengthen the semi-mature state of the former group (Fig. 2C). Remarkably, this common signature is also completely shared Doxorubicin chemical structure among the stimuli TNF, mfVSG, and MiTat1.5 sVSG, since no different or additional genes were induced (triple field with zero genes, Fig. 2B). Thus, the semi-mature DC signature was represented by upregulation of CD40, CD72, IL-1α, IL-1β, IL-6, CXCL2,
SOCS3, Jagged-1, Pleckstrin-2 (Plek2),
serum amyloid 3(Saa3), ladinin (Lad), follistatin, (FST), activin (Inhba), and downregulation of PGE-receptor 3 (Ptger3), CD62L (Sell) SIGNR2 (CD209c). In contrast, the fully matured DC signature of genes induced by LPS include the common 24 genes, but regulated additional 4498 genes that were not shared with the other stimuli (Fig. 2B). The exclusive gene signatures induced by TNF alone (Supporting Information Fig. 2) or the comparisons of mfVSG with MiTat1.5 sVSG (Supporting Information Fig. 3) were not marked by a strong immunological signature of gene regulation. Taken together, the common signature of DCs matured by TNF, mfVSG, and MiTat1.5 sVSG induces far fewer genes than LPS, which are mainly characterized by a common signature of 24 mostly inflammatory genes. To dissect Galunisertib datasheet over the importance of the partial DC maturation phenotypes in directing distinct Th cell differentiation patterns, we cocultured DCs with OVA-specific TCR-transgenic CD4+ OT-II T cells and checked the Th-cell profile by intracellular cytokine staining. Polarizing by LPS showed a clear shift toward IFN-γ, indicating a Th1-cell profile. DC maturation with TNF and mfVSG shifted the T cells toward a Th2/Th9-cell pattern
and DC stimulation with MiTat1.5 sVSG heavily reduced Th2-cell and Th9-cell but left the Th1-cell background profile unaltered (Fig. 3A and B and Supporting Information Fig. 4). Furthermore, induction of IL-17 production in T cells was negligible under all conditions (Supporting Information Fig. 4) and T cells did not produce anti-inflammatory IL-10 after one round of DC stimulation (data not shown) and as we reported previously 23. Earlier reports demonstrated that BM-derived DCs efficiently induced CD4+CD25+FoxP3+ Treg cells in vitro predominately in the presence of exogenously supplied TGF-β 41. Indeed, hardly any Treg-cell generation could be detected in the absence of exogenous TGF-β irrespective of the maturation phenotype of the DCs (Supporting Information Fig. 5A). Nevertheless, DCs matured with TNF, mfVSG, or MiTat1.