使用质谱流式细胞仪（CyTOF）进行抗原特异性T细胞的质谱流式细胞术检测。
Mass Cytometry Assays for Antigen-Specific T Cells Using CyTOF.
作者信息
Lin Dongxia, Maecker Holden T
机构信息
Fluidigm Corporation, South San Francisco, CA, 94080, USA.
Human Immune Monitoring Center, Institute for Immunity, Transplantation, and Infection, Stanford University School of Medicine, Fairchild Science Building, 299 Campus Drive, Stanford, CA, 94305-5124, USA.
出版信息
Methods Mol Biol. 2018;1678:37-47. doi: 10.1007/978-1-4939-7346-0_3.
Abstract
T Cells specific for a single antigen tend to be rare, even after expansion of memory cells. They are commonly detected by in vitro stimulation with peptides or protein, followed by staining for intracellular cytokines. In this protocol, we use CyTOF mass cytometry to collect single-cell data on a large number of cytokines/chemokines, as well as cell-surface proteins that characterize T cells and other immune cells. We also include a method for magnetic bead enrichment of antigen-stimulated T cells, based on their expression of CD154 and CD69. Coupling magnetic enrichment with highly multi-parameter mass cytometry, this method enables the ability to dissect the frequency, phenotype, and function of antigen-specific T cells in greater detail than previously possible. Rare cell subsets can be examined, while minimizing run times on the CyTOF.
摘要
即使在记忆细胞扩增后，针对单一抗原的T细胞往往也很稀少。通常通过用肽或蛋白质进行体外刺激，然后对细胞内细胞因子进行染色来检测它们。在本方案中，我们使用质谱流式细胞术（CyTOF）来收集大量细胞因子/趋化因子以及表征T细胞和其他免疫细胞的细胞表面蛋白的单细胞数据。我们还包括一种基于抗原刺激的T细胞对CD154和CD69的表达进行磁珠富集的方法。将磁富集与高度多参数的质谱流式细胞术相结合，该方法能够比以往更详细地剖析抗原特异性T细胞的频率、表型和功能。可以检测稀有细胞亚群，同时将在CyTOF上的运行时间降至最低。
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