Hennies Cassandra M, Lehn Maria A, Janssen Edith M
Division of Immunobiology, Cincinnati Children's Hospital Medical Center and the University of Cincinnati College of Medicine, Cincinnati, OH, USA.
Division of Immunobiology, Cincinnati Children's Hospital Medical Center and the University of Cincinnati College of Medicine, Cincinnati, OH, USA.
J Immunol Methods. 2015 Aug;423:18-28. doi: 10.1016/j.jim.2015.04.023. Epub 2015 May 9.
Presentation of antigenic peptides in MHC class II (MHCII) on dendritic cells (DCs) is the first step in the activation of antigen-specific CD4(+)T cells. The expression of surface MHCII-peptide complexes is tightly regulated as the frequency of MHCII-peptide complexes can affect the magnitude, as well as the phenotype of the ensuing CD4(+)T cell response. The surface MHCII-peptide levels are determined by the balance between expression of newly generated complexes, complex internalization, and their subsequent re-emergence or degradation. However, the molecular mechanisms that underpin these processes are still poorly understood. Here we describe a multispectral imaging flow cytometry assay to visualize MHCII trafficking that can be used as a tool to dissect the molecular mechanisms that regulate MHCII homeostasis in primary mouse and human DCs.
树突状细胞（DC）上的主要组织相容性复合体II类（MHCII）对抗原肽的呈递是激活抗原特异性CD4(+)T细胞的第一步。表面MHCII-肽复合物的表达受到严格调控，因为MHCII-肽复合物的频率会影响随后CD4(+)T细胞反应的强度和表型。表面MHCII-肽水平由新生成复合物的表达、复合物内化及其随后的重新出现或降解之间的平衡决定。然而，支撑这些过程的分子机制仍知之甚少。在这里，我们描述了一种多光谱成像流式细胞术检测方法，用于可视化MHCII的运输，该方法可作为一种工具，用于剖析调节原代小鼠和人DC中MHCII稳态的分子机制。
