Dendritic cell subsets in autoimmune diabetes

利用fms样酪氨酸激酶3配体（Flt3L）刺激骨髓细胞培养，我们发现小鼠和人类自身免疫性糖尿病易感基因IL-2抑制DC的发展。IL-2可能在MDP阶段起作用，因为它们是表达IL-2Ra的前体，而MDPs在Flt3L和IL-2的培养中积累。我们现在发现，当将IL2添加到Flt3L BMDC培养物中时，单核细胞和巨噬细胞的前体增加，这表明IL2可以将DC的发育转变为单核细胞谱系。我们测量了在IL-2存在下发展的dc中基因表达的变化，发现编码bcl2l11的基因与IL-2一起升高，CD135 （flt3）与IL-2一起降低。这两种变化在蛋白质水平上得到证实。此外，我们发现在IL-2存在下发育的DC在Flt3L刺激下显示出较低的磷酸化stat3，这可能解释了DC发育的阻滞。

Using cultures of bone marrow cells stimulated with FMS-like tyrosine kinase 3 ligand (Flt3L), we have shown that IL-2, an autoimmune diabetes susceptibility gene in both mouse and human, inhibits DC development. IL-2 is likely acting at the MDP stage because those are the precursors that express the IL-2Ra, and MDPs accumulate in cultures with Flt3L and IL-2. We now find that when IL2 is added to Flt3L BMDC cultures, the precursor of monocytes and macrophages increases, suggesting that IL2 can shift development from the DC to monocyte lineage. We measured gene expression changes in DCs that develop in the presence of IL-2 and found the bim-encoding gene bcl2l11 up with IL-2 and CD135 (flt3) down with IL-2. Both of these changes were confirmed at the protein level. Furthermore, we showed that DCs that develop in the presence of IL-2 display lower phospho-STAT3 in response to Flt3L stimulation, which may explain the block in DC development. We are also interested in determining how DCs differ in NOD mice compared to non-autoimmune strains, and how DCs are different in NOD mice at different diabetes pathogenesis states and disease sites. We have compared responses of 8-10 week old NOD and B6 spleen and lymph node DCs to TLRL which gives us information about both baseline and stimulated activity of DCs in mice with chronic autoimmunity but before overt disease initiates. Readouts include gene expression analysis, flow cytometry and elisa. We have identified differences in the cytokine and costimulatory responses of NOD DCs. We also find that IFN-responsive genes are induced less in NOD DCs after CpG stimulation, and are following up on this to determine what pathways are responsible for this altered response, as well as what the consequences are for DC-T cell interactions and diabetes pathogenesis.