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.
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。
