Antigen processing in the secretory pathway

描述（由申请人提供）：我们研究的长期目标是了解和操纵在细胞表面产生数千种肽/MHC I类复合物的抗原加工机制。与描述抗原肽仅在细胞质中产生的教科书模型相反，最近的研究结果表明抗原加工发生在细胞质中并且在分泌途径中特别是内质网（ER）中继续。介导抗原肽修剪的蛋白酶最近已被鉴定，并被称为ERAAP，用于与抗原加工相关的ER氨肽酶。 在这里，我们建议填补我们的理解肽修剪如何发生在ER及其对CD 8 T细胞库在生理背景下的影响的差距。我们将使用我们在实验室开发的独特工具进行分析。这些包括（a）ERAAP敲除小鼠，和（B）用于分析抗原加工途径的最终输出以及在正常和专职抗原呈递细胞中产生的难以捉摸的蛋白水解中间体的新方法。

DESCRIPTION (provided by applicant): The long-term goal of our research is to understand and manipulate the antigen processing mechanisms that yields thousands of peptide/MHC class I complexes on the cell surface. Contrary to the text book model which depicts the antigenic peptides being generated solely in the cytoplasm, recent findings indicate that antigen processing occurs both in the cytoplasm and continues in the secretory pathway specifically the endoplasmic reticulum (ER). The protease that mediates the trimming of antigenic peptides has recently been identified and termed ERAAP, for an ER aminopeptidase associated with antigen processing. Here we propose to fill the gaps in our understanding of how peptide trimming occurs in the ER and its impact on the CD8 T cell repertoire in a physiological context. We will use the unique tools we have developed in our laboratory for this analysis. These include (a) the ERAAP knock-out mice, and (b) novel methods for analysis of final output of the antigen processing pathway as well as the elusive proteolytic intermediates that are generated in normal and professional antigen presenting cells.