Antigen processing in the secretory pathway

描述(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个肽/MHC I类复合体的抗原处理机制。与仅在细胞质中产生抗原肽的教科书模型相反，最近的发现表明，抗原处理既发生在细胞质中，又继续在分泌途径中，特别是内质网(ER)。最近发现了一种调节抗原肽修剪的蛋白酶，命名为ERAAP，是一种与抗原加工相关的内质网氨基肽酶。 在这里，我们建议填补我们在理解内质网中如何发生肽修剪及其在生理背景下对CD8T细胞库的影响方面的空白。我们将使用我们实验室开发的独特工具进行这一分析。这些方法包括(A)ERAAP基因敲除小鼠，以及(B)分析抗原处理途径的最终输出以及在正常和专业抗原提呈细胞中产生的难以捉摸的蛋白分解中间体的新方法。

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.