Using dendritic cells to present beta cell antigens in vivo for manipulation of diabetes pathogenesis

为了寻找DC靶向胰岛素肽改变糖尿病发病机制的条件，我们建立了一系列的工具和小鼠模型。我们已经创造和生产了嵌合抗体，可以针对DEC205+和DCIR2+DC的几种胰岛素抗原。此外，我们还优化了MHC II类I-A-G7四聚体与胰岛素肽的染色，以识别体内胰岛素特异性的CD4T细胞，并改进了我们的胰腺和胰岛分离，以识别浸润性T细胞。我们还在实验室建立了几个小鼠模型，包括胰岛素2基因敲除和杂合子小鼠。小鼠有两个胰岛素基因，因为只有Ins2在胸腺表达，消除INS2的一个或两个等位基因会减少对胰岛素特异性T细胞的负选择，导致糖尿病加速，T细胞和B细胞的反应更多地集中在胰岛素上。我们现在正在测试针对几个DC亚群的胰岛素多肽嵌合抗体如何影响内源性胰岛素特异性CD4T细胞反应和糖尿病的发病机制(包括胰岛素炎和高血糖)。

In order to find conditions in which DC-targeted insulin peptide can alter diabetes pathogenesis, we have set up a series of tools and mouse models. We have created and produced chimeric antibodies that can target several insulin antigens to both DEC205+ and DCIR2+ DCs. In addition, we have optimized staining of MHC class II I-A-g7 tetramers with an insulin peptide for identifying insulin-specific CD4 T cells in vivo, and improved our pancreas and islet isolation to identify infiltrating T cells. We have also set up several mouse models in our lab, including the Insulin 2 knockout and heterozygous mice. Mice have 2 insulin genes, and because only Ins2 is expressed in the thymus, elimination of one or both alleles of ins2 reduces negative selection for insulin-specific T cells, resulting in accelerated diabetes in which the T and B cell responses are more insulin focused. We are now testing how chimeric antibodies that target insulin peptide to several DC subsets affects endogenous insulin-specific CD4 T cell responses, and diabetes pathogenesis (both insulitis and hyperglycemia).