使用 II 类 MHC 四聚体染色和富集技术鉴定人类抗原特异性 T 细胞。
Identification of Human Antigen-Specific T Cells Using Class II MHC Tetramer Staining and Enrichment.
机构信息
Department of Medicine, Division of Rheumatology, Perelman School of Medicine, Institute for Immunology, University of Pennsylvania, Philadelphia, PA, USA.
Corporal Michael J Crescenz VA Medical Center, Philadelphia, PA, USA.
出版信息
Methods Mol Biol. 2022;2574:31-40. doi: 10.1007/978-1-0716-2712-9_3.
The development of peptide-major histocompatibility complex tetramers has enabled direct characterization and enumeration of antigen-specific T cells. However, the weaker interaction between class II tetramers and CD4 T cells increases the challenge of using tetramers to analyze CD4 T cell responses. Here, we provide an optimized class II tetramer staining protocol with a magnetic-bead enrichment strategy for the detection and functional analyses of human antigen-specific CD4 T cells. This approach enables direct sampling of lymphocytes that recognize specific peptide-MHC complexes, including rare pathogen-specific CD4 T cells from unexposed individuals.
肽-主要组织相容性复合物四聚体的发展使抗原特异性 T 细胞的直接鉴定和计数成为可能。然而，II 类四聚体与 CD4 T 细胞之间较弱的相互作用增加了使用四聚体分析 CD4 T 细胞反应的挑战。在这里，我们提供了一种优化的 II 类四聚体染色方案，结合磁珠富集策略，用于检测和分析人类抗原特异性 CD4 T 细胞的功能。这种方法能够直接采集识别特定肽-MHC 复合物的淋巴细胞，包括来自未接触个体的稀有病原体特异性 CD4 T 细胞。
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