Analyzing the function of the endocytic receptor DEC205 in vitro and in vivo during delivery and presentation of antigens in brain endothelial cells.

分析内吞受体 DEC205 在体外和体内在脑内皮细胞中递送和呈递抗原过程中的功能。

• 批准号: 459618779
• 负责人: Professor Dr. Karsten Mahnke
• 金额: --
• 依托单位: Universitäts-Hautklinik
• 依托单位国家: 德国
• 项目类别: Research Grants
• 资助国家: 德国
• 项目状态: 未结题
• 来源: https://gepris.dfg.de/gepris/projekt/459618779?language=en
• 关键词: Analyzing; function; endocytic; receptor; DEC205

The antigen uptake receptor DEC205 was originally described in dendritic cells (DCs). Within the intracellular domain we characterized three defined sequences, mediating (a) the uptake of ligands, (b) the targeting to deeper endosomal compartments and (c) the recycling back to the surface of DCs. This makes DEC205 a highly specialized receptor for absorptive endocytosis. Beyond DCs, DEC205 is exclusively expressed by endothelial cells (ECs) of the brain. While ECs are not classical antigen presenting cells, our preliminary data show that DEC205 mediates uptake of ligands into ECs, followed by presentation to T cells. Thus, DEC205 may serve as “linker” molecule connecting the function of ECs with those of the immune system. To analyze the functions of DEC205 in ECs in detail, we will introduce targeted mutations into its domains and follow the intracellular routing of DEC205 and its ligands through endosomal compartments. Possible adapter molecules of DEC205, i.e. members of the endosomal sorting complexes will be analyzed by co-labeling and immunoprecipitation with anti-DEC205 antibodies. For in vivo investigations of normal and inflamed brain endothelia, we will make use of the experimental allergic encephalomyelitis (EAE) model. We will couple surrogate ligands to anti-DEC205 antibodies and target brain ECs in normal and EAE induced mice. Thereafter, we will analyze the antigen uptake by ECs, the presentation of antigens to T cells, as well as their differentiation (i.e. Treg, Th1, Th2, anergy) during EAE or healthy conditions. In summary, we will define the functions of the DEC205 receptor at the interface of the vascular endothelium and the immune system.

抗原摄取受体DEC205最初是在树突状细胞（DC）中描述的。在细胞内结构域中，我们表征了三个定义的序列，介导（a）配体的摄取，（b）靶向更深的内体室的靶向，以及（c）回收回到DC的表面。这使DEC205成为吸收性内吞作用的高度专业化受体。除了DC之外，DEC205仅由大脑的内皮细胞（EC）表达。尽管EC不是经典的抗原呈现细胞，但我们的初步数据表明，DEC205介导了配体对EC的摄取，然后将其介绍给T细胞。这是DEC205可以用作“接头”分子，将EC的功能与免疫系统的功能联系起来。为了详细分析DEC205的功能，我们将将靶向突变引入其域，并遵循DEC205及其配体的细胞内路由，通过内体隔室。 DEC205的可能的衔接分子，即内体分选复合物的成员将通过与抗DEC205抗体共同标记和免疫沉淀来分析。为了对正常和发炎的脑内皮进行体内研究，我们将利用实验性过敏性脑脊髓炎（EAE）模型。我们将在正常和EAE诱导的小鼠中将替代配体与抗DEC205抗体和靶向脑EC靶向。此后，我们将分析EC的抗原摄取，在EAE或健康状况下，抗原向T细胞的呈现及其分化（即Treg，Th1，Th2，Anergy）。总之，我们将在血管内皮和免疫系统的界面上定义DEC205受体的功能。