Autophagy-mediated antigen presentation for CD1d-restricted NKT cell recognition

在拟议的研究中，我们的目的是研究自噬是否通过将抗原（Ags）递送到cd1装载室来调节自然杀伤T （NKT）细胞反应。自噬是一种高度保守的溶酶体分解代谢途径，在此过程中，细胞器和长寿命蛋白质被降解，以提供细胞生存所需的营养。除了参与体内平衡、细胞死亡和细胞清除外，自噬似乎在抗原呈递细胞（APCs）中通过自噬体促进抗原呈递方面起着至关重要的作用。NKT细胞是一种T淋巴细胞亚群，与传统T细胞和自然杀伤细胞具有相同的功能特征。NKT细胞通过识别CD1d分子呈递的脂质Ags，被认为在先天免疫和适应性免疫过程的中介中起关键作用。我们假设自噬通过脂质Ags转运到cd1装载室促进了NKT细胞的反应。为了解决上述假设，我们将分析在稳态和免疫激活后，专业apc中自噬体是否与cd1装载腔室共定位，以及apc中自噬的破坏是否会减少CD1d的呈递和NKT细胞的激活。用于进行所需分析的实验设置将包括流式细胞术、人类和小鼠骨髓细胞系、ELISA、sirna介导的敲除、小鼠实验，包括条件敲除小鼠以破坏自噬，以及通过共聚焦显微镜进行免疫细胞化学共定位研究。这些结果有望进一步加深我们对自噬介导的银通过CD1d分子呈递和连续的NKT细胞反应之间的分子相互作用的理解。调节cd1限制性NKT细胞活性可能为治疗和预防（自身）免疫介导的疾病提供新的治疗选择

In the proposed research, we aim to investigate whether autophagy regulates natural killer T (NKT) cell responses through delivery of antigens (Ags) to CD1d-loading compartments. Autophagy is a highly conserved lysosomal catabolic pathway during which cell organelles and long-lived proteins are degraded in order to provide nutrients for cell survival. Aside from its involvement in homeostasis, cell death and cellular clearance, autophagy appears to play a crucial role in facilitating antigen presentation via autophagosomes in antigen-presenting cells (APCs). NKT cells represent a T lymphocyte subpopulation sharing functional characteristics with both conventional T cells as well as natural killer cells. NKT cells are believed to be pivotally involved in the mediation between innate and adaptive immune processes by recognizing lipid Ags presented via CD1d molecules. We hypothesize that autophagy facilitates NKT cell responses through transport of lipid Ags to CD1d-loading compartments. To address the aforementioned hypothesis, we will analyze whether autophagosomes colocalize with CD1d-loading compartments in professional APCs during the steady state and following immune activation as well as whether disruption of autophagy in APCs reduces CD1d presentation and NKT cell activation in vitro and in vivo. The experimental set up used to carry out the required analyses will include flow cytometry, human and mouse myeloid cell lines, ELISA, siRNA-mediated knockdown, mice experiments involving conditional knockout mice for disruption of autophagy as well as immunocytochemical co-localization studies via confocal microscopy. The results are expected to further our understanding of the molecular interactions between autophagy-mediated Ag presentation via CD1d molecules and consecutive NKT cell responses. Modulation of CD1d-restricted NKT cell activity might provide new therapeutic options for the treatment and prevention of (auto-) immune-mediated conditions