多重MHC I类四聚体与质谱流式细胞术的核内染色相结合

Multiplex MHC Class I Tetramer Combined with Intranuclear Staining by Mass Cytometry.

作者信息

Simoni Yannick, Fehlings Michael, Newell Evan W

机构信息

Agency for Science, Technology and Research (A*STAR), Singapore Immunology Network (SIgN), Singapore, Singapore.

Fred Hutch Cancer Research Center, Vaccine and Infectious Disease Division, Seattle, WA, USA.

出版信息

Methods Mol Biol. 2019;1989:147-158. doi: 10.1007/978-1-4939-9454-0_11.

DOI:10.1007/978-1-4939-9454-0_11

PMID:31077105

Abstract

Antigen-specific CD8 T cells play a crucial role in the host protective immune response against viruses, tumors, and other diseases. Major histocompatibility complex (MHC) class I tetramers allow for a direct detection of such antigen-specific CD8 T cells. Using mass cytometry together with multiplex MHC class I tetramer staining, we are able to screen more than 1000 different antigen candidates simultaneously across tissues in health and disease, while retaining the possibility to deliver an in-depth characterization of antigen-specific CD8 T cells and associated phenotypes. Here we describe the method for a MHC class I tetramer multiplexing approach together with intracellular antibody staining for a parallel phenotypic cell characterization using mass cytometry in human specimens.

摘要

抗原特异性CD8 T细胞在宿主针对病毒、肿瘤和其他疾病的保护性免疫反应中发挥关键作用。主要组织相容性复合体（MHC）I类四聚体能够直接检测此类抗原特异性CD8 T细胞。通过将质谱流式细胞术与多重MHC I类四聚体染色相结合，我们能够在健康和疾病状态下同时对超过1000种不同的抗原候选物进行跨组织筛选，同时还能够深入表征抗原特异性CD8 T细胞及其相关表型。在此，我们描述了一种用于MHC I类四聚体多重分析方法以及细胞内抗体染色的方法，以便使用质谱流式细胞术对人类样本中的细胞进行平行表型特征分析。