脱颗粒
细胞毒性T细胞
生物
分子生物学
细胞溶解
细胞生物学
抗原
细胞内
化学
抗原提呈细胞
CD28
CD8型
T细胞
穿孔素
免疫学
受体
免疫系统
生物化学
体外
作者
Michael R. Betts,Jason M. Brenchley,David A. Price,Stephen C. De Rosa,Daniel C. Douek,Mario Roederer,Richard A. Koup
标识
DOI:10.1016/s0022-1759(03)00265-5
摘要
Flow cytometric detection of antigen-specific CD8+ T cells has previously been limited to MHC-class I tetramer staining or intracellular cytokine production, neither of which measure the cytolytic potential of these cells. Here we present a novel technique to enumerate antigen-specific CD8+ T cells using a marker expressed on the cell surface following activation induced degranulation, a necessary precursor of cytolysis. This assay measures the exposure of CD107a and b, present in the membrane of cytotoxic granules, onto the cell surface as a result of degranulation. Acquisition of cell surface CD107a and b is associated with loss of intracellular perforin and is inhibited by colchicine, indicating that exposure of CD107a and b to the cell surface is dependent on degranulation. CD107a and b are expressed on the cell surface of CD8+ T cells following activation with cognate peptide, concordant with production of intracellular IFNγ. Finally, CD107-expressing CD8+ T cells are shown to mediate cytolytic activity in an antigen-specific manner. Measurement of CD107a and b expression can also be combined with MHC-class I tetramer labeling and intracellular cytokine staining to provide a more complete assessment of the functionality of CD8+T cells expressing cognate T cell receptors (TCR).
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