If DCs were the primary APC for priming naïve Th cells in EAE, an increased naïve Th-cell compartment after DC depletion would be expected. Thus, our data argues for that another cell type is the primary APC for priming naïve Th cells to become autoimmune. Differentiation of Th17 or Th1 cells was also not affected by the DC depletion. Since we have previously shown that pDCs regulate the Th17 response toward MOG in EAE , we tested whether pDCs were also depleted in CD11c-DTR and bone marrow chimeras after DTx treatment. Two different flow cytometry methods clearly showed that pDCs were not depleted by the DTx injection.
To further examine the role of DCs on Th differentiation, DC maturation and Treg-cell responses were studied. DC maturation 10 days after MOG immunization was not impaired after DC ablation a day before EAE induction. We have Ulixertinib mouse previously shown that IL-6 and IL-23p40 expression is upregulated in mDCs by a MyD88-dependent mechanism in EAE . Another possiblity was that Treg cells were affected by the DC depletion and subsequently ameliorated the EAE severity. The number of Treg cells in the spleen was however not affected by the DC depletion. Adriamycin order After constituitive ablation of DCs, Treg-cell numbers
are reduced [9, 10]. The difference between our data and their systems is probably caused by the short ablation period and the fact that thymic selection prior to DTx injection is most likely not affected in our system. Others have clearly demonstrated that DCs reactivate primed encephalitogenic Th cells in the CNS during development of EAE . In their system, the myelin-reactive Th cells were however transferred to the mice after priming. In an EAE model of epitope spreading, naïve Th cells reactive to proteolipid protein139–151 were primed probably by DCs in the CNS . An ongoing myelin-reactive Th-cell response was required for epitope spreading to occur. The infiltration of DCs into the CNS was not affected in our transient
system, and we focused on priming and de novo differentiation of naïve Th cells to become myelin-reactive, where DCs appear to have no major role Inositol monophosphatase 1 or are redundant. A reduced or an abolished CD11c expression on DCs during the development of EAE could have rendered the CD11c-DTR mice and bone marrow chimeras resistant to the DC depletion and skewed our results. We have however previously observed similar numbers of CD11chi MHC II+ mDCs in the spleen during sorting of mDC at 4 and 10 days after MOG immunization and in unimmunized mice  (A. Lobell, unpublished observations). It is therefore unlikely that reduced CD11c expression explains the observed phenotype. Unexpectedly, transient ablation of DCs before or after EAE induction does not affect priming of Th cells or de novo differentiation of autoimmune, MOG-induced Th17 and Th1-cell responses.