From the results presented here, one could expect Pds5B mutations to be causative of this disease

icle. The FITChi+ and FITC CD11c+ populations derived from the dLNs were isolated by FACS sorting. The percentage of each cell phenotype is listed in the respective quadrant. Approximately 5% of the FITChi+ cells became FITClo+ compared with the cell-sorted populations immediately after sorting. These DC apparently lost FITC taken up during the FACs staining step or fixation in PFA. A representative experiment from an n = 2 is shown. was applied to dorsal skin, as opposed to OVAFITC which gave a very weak response similar to observations of other groups, we detected a slight but significant decrease in the number of FITChi+ CD11c+ DC that migrated from the skin to the dLN in Compound A-treated mice. This difference was not detected in earlier studies using CRTH2/ mice, as the percentage of FITC+ cells was compared with whole lymph node cell population, as opposed to just the CD11c+ cells. Further, our results show that a high percentage of the FITChi+ cells in the dLN express CD11c, strongly suggesting that these cells migrated from the skin, as opposed to acquiring FITC via the lymph or blood stream. Co-culturing of the different populations of CD11c+ DC, those expressing high levels of FITC, the resident CD11c+ DC in the lymph nodes that were FITC and splenic-derived CD11c+ DC, showed striking effects on their ability to activate naive T cells. As noted previously, FITChi+ CD11c+ DC elicited a much greater A-83-01 site IL-17A response compared with splenic-derived CD11c+ DC co-cultured with naive T cells. However, the FITChi+ CD11c+ DC isolated from Compound A-treated mice elicited substantially decreased levels of IL-17A, IFN-c, IL-10 and IL-6 upon co-culture with naive DO11.10 TCR transgenic T cells compared with vehicle-treated animals. As IL-6 has been shown to play a role in the differentiation of Th17 cells, perhaps this decrease in IL-17A may be linked to the decrease of IL-6 produced by naive T cells co-cultured with FITChi+ CD11c+ DC from Compound A-treated mice. However, this decrease is already detected in cultures after 24 h and the CD11c+ cells do not make any IL-6 when cultured in the absence of T cells or antigen. Additionally, no significant differences in the amount of TGF-b cytokine or IL-23 mRNA levels were detected between the various DC populations. This decreased amount of cytokine production does not appear to be mediated by the induction of a regulatory T cell population, as we found no increase of FoxP3+ cells, or CTLA-4+ cells in the cultures after 72 h. The levels of IL-10 were not increased in the cultures containing the FITChi+ CD11c+ DC from Compound A-treated mice. IL-6 was present in the same cultures and if a Treg population was induced, IL-6 levels would be expected to be greatly decreased. The FITChi+ DC derived from Compound A-treated mice did not appear to induce anergy in the naive T cell population, as the levels of IL-4 produced were not significantly different among the different DC supernatant was removed at 72 h. The culture PubMed ID:http://www.ncbi.nlm.nih.gov/pubmed/19825521 supernatants were assayed by ELISA for IL-17A, IFN-c, IL-10, IL-6 and IL-4 levels, and additionally the IL-17A and IFN-c levels were determined for the 24 and 48 h-time points. The results from the co-cultures containing the dLN CD11c+ DC are labeled dLN CD11c+ DC. Each bar graph shows a representative experiment of a minimum of three independent experiments. Each column represents the mean cytokine value obtained from that experiment 6 SEM. For the kinetic analysis of IL-17A and IFN-