For 5 ng/mL IL-17 (**P =

For 5 ng/mL IL-17 (**P = <0.01), and to 42 for 50 ng/mL IL-
For 5 ng/mL IL-17 (**P = <0.01), and to 42 for 50 ng/mL IL-17 (**P = <0.01), each compared with control stimulations. (F) Internodal distances after exposure to IL-17 revealed a significant decrease with 5 ng/mL IL-17 (*P = <0.05) and 50 ng/mL IL-17 (*P = <0.05). Analysis of variances between the independent experiments revealed no significant difference. (G) Myelin quantification of Sudan-stained DRG co-cultures after treatment with IL-17, an IL-17-neutralizing antibody (IL-17ab), and co-stimulation of three independent experiments, respectively. The myelin-inhibitory effect of IL-17 was reduced by 81 after supplementation with IL-17ab. Myelination was restored to the base level, similar to treatment with the antibody alone.Accordingly, the internodal distance, a parameter for quantitative myelin synthesis, was reduced Aviptadil price significantly following stimulation with 5 and 50 ng/mL IL-17 (Figure 2F). Measurement of fiber diameter in Sudan stained cultures did not reveal a significant difference for the IL-17-treated cultures (data not shown). Costimulation of DRG co-cultures with IL-17 and an IL17-neutralizing antibody reduced the myelin inhibitory effect by about 81 (Figure 2G). mRNA expression of genes associated with myelination of rSCs was analyzed after stimulation with IL-17 for 10 days. The mRNA expression of KROX-20 decreased significantly with 5 and 50 ng/mL IL-17 (Figure 3A). PubMed ID:https://www.ncbi.nlm.nih.gov/pubmed/28404814 For P0, the mRNA expression decreased significantly with 5 ng/mL IL-17 and decreased non-significantly with 50 ng/mL IL-17 (Figure 3A). Evaluation of myelin morphology using electron microscopy did not reveal any morphological alterations when compared with control cultures (Figure 3B).IL-17 does not impede neuronal outgrowth and cell survivalpresentation, MHCI and MHCII, as well as TAPII, were analyzed using immunocytochemistry and quantified using densitometry. SCs displayed expression of all three molecules: MHCI > TAPII > MHCII. MHCI expression was significantly increased in a dose-dependent manner after treatment with IL-17 for 72 h (Figure 5A). Compared to MHCI, MHCII exhibited a fainter expression under control conditions; its expression was slightly but not significantly increased after 50 ng/mL IL-17, but revealed a significant increase at a concentration of 0.5 ng/ mL IL-17 (Figure 5B). The expression of TAPII was slightly, though not significantly, increased in a dose-dependent manner after stimulation with IL-17 (Figure 5C).To exclude a direct effect of IL-17 on neuronal outgrowth and a consecutive effect on myelin synthesis, we performed neurofilament (NF) staining and quantification in mouse DRG co-cultures after exposure to IL-17 for 21 days. Neuronal fibers did not show a significant reduction in NF after IL-17 exposure using densitometry analysis, normalized to background fluorescence (Figure 4A, B). To further exclude a direct effect of IL-17 on SC survival, viability assays were performed in rSCs. Stimulation with 0.5 to 50 ng/mL IL-17 did not cause any significant alterations in SC viability (Figure 4C). To analyze additional mediators of inflammation and myelination, the activity of MMPs was assessed. IL-17treated DRG co-culture supernatants revealed a significant dose-dependent downregulation of MMP-2 activity and a dose-dependent upregulation of MMP-9 activity, assessed in gelatine zymography and quantified using densitometry (Figure 4D, E).IL-17 increases major histocompatibility complex and transporter associated with antigen presenta.