Keetz K, Knapp M, Schulze Zur Wiesch J
Infectious Diseases Unit, I. Department of Medicine, University Medical Center Hamburg-Eppendorf, Hamburg, Germany.
Methods Mol Biol. 2025;2904:193-209. doi: 10.1007/978-1-0716-4414-0_14.
This chapter presents a detailed protocol for peptide-major histocompatibility complex (pMHC) multimer staining and magnetic activated cell separation, offering a step-by-step guide to isolating and studying low-frequency antigen-specific T cells. This methodological approach provides a means to evaluate the phenotype and function of antigen-specific T cells, potentially facilitating the assessment of novel therapeutic interventions and aiding in understanding T cell exhaustion in chronic viral diseases. Despite challenges such as the necessity of knowing the epitope and HLA restrictions as well as the intricate nature of the pMHC multimer staining process, this protocol serves as a fundamental tool for both basic research and translational applications in chronic viral infections.
本章介绍了肽-主要组织相容性复合体(pMHC)多聚体染色和磁珠激活细胞分选的详细方案,提供了分离和研究低频抗原特异性T细胞的分步指南。这种方法提供了一种评估抗原特异性T细胞表型和功能的手段,有可能促进对新型治疗干预措施的评估,并有助于理解慢性病毒性疾病中的T细胞耗竭。尽管存在诸如需要了解表位和HLA限制以及pMHC多聚体染色过程的复杂性等挑战,但该方案仍是慢性病毒感染基础研究和转化应用的基本工具。