Dendritic cell subsets in autoimmune diabetes

使用FMS样酪氨酸激酶3配体（Flt 3L）刺激的骨髓细胞培养物，我们已经表明，IL-2，在小鼠和人类中的自身免疫性糖尿病易感基因，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-2 Ra的前体，并且MDP在具有Flt 3L和IL-2的培养物中积累。此外，我们发现当IL 2加入Flt 3L BMDC培养物中时，单核细胞和巨噬细胞的前体增加，表明IL 2可以将DC的发育转变为单核细胞谱系。 我们现在有数据显示，在IL-2存在下，糖尿病易感NOD小鼠与糖尿病抗性B6对照小鼠的DC基因表达变化。 我们还感兴趣的是，确定NOD小鼠中的DC与非自身免疫菌株相比如何不同，以及NOD小鼠中的DC在不同的糖尿病发病机制状态和疾病部位如何不同。 我们开始比较8-10周龄NOD和B6脾和淋巴结DC对TLRL的应答。 这为我们提供了关于慢性自身免疫小鼠中DC的基线和刺激活性的信息，但在明显的疾病开始之前。 读数包括基因表达分析，流式细胞术和elisa。我们正在发现NOD DC的细胞因子和共刺激反应的差异，现在正在确定哪些信号通路可能参与这些改变的反应。

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. In addition, we 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 now have data showing gene expression changes in DCs that develop in the presence of IL-2, comparing both the diabetes susceptible NOD mice to diabetes resistant B6 control mice. 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 started by comparing responses of 8-10 week old NOD and B6 spleen and lymph node DCs to TLRL. This 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 are finding differences in the cytokine and costimulatory responses of NOD DCs and are now determining what signaling pathways may be involved in these altered responses.