小鼠CD8 T细胞杀伤试验。
Murine CD8 T Cell Killing Assay.
作者信息
Kim Myoungjoo V, Ouyang Weiming, Liao Will, Zhang Michael Q, Li Ming O
机构信息
Department of Immunobiology, Yale University School of Medicine, New Haven, USA.
Center for Drug Evaluation and Research, U.S. Food and Drug Administration, Bethesda, USA.
出版信息
Bio Protoc. 2014 Jul 5;4(13). doi: 10.21769/BioProtoc.1172.
Antigen-specific killing ability of effector CD8 T cells is critical for protective immunity against infection. Here, we describe cytotoxic T cell assay to examine effector function of antigen-specific CD8 T cells. Mice infected with Listeria monocytogenes () expressing chicken ovalbumin as a model antigen mount ovalbumin-specific CD8 T cell responses. Effector CD8 T cell function is determined by mixed transfer of OVA peptide-pulsed target cells with control target cells into the previously immunized mice. Difference in CFSE expression levels clearly marks two distinct populations: Antigen-pulsed target cells-CFSE vs. unpulsed target cells-CFSE. The frequencies between antigen-pulsed target cells and control target cells are used as readouts of antigen-specific killing.
效应性CD8 T细胞的抗原特异性杀伤能力对于抗感染的保护性免疫至关重要。在此，我们描述了一种细胞毒性T细胞检测方法，用于检测抗原特异性CD8 T细胞的效应功能。以表达鸡卵清蛋白作为模型抗原的单核细胞增生李斯特菌感染的小鼠会产生卵清蛋白特异性CD8 T细胞应答。效应性CD8 T细胞功能通过将卵清蛋白肽脉冲处理的靶细胞与对照靶细胞混合转移到预先免疫的小鼠中来确定。CFSE表达水平的差异清楚地标记了两个不同的群体：抗原脉冲处理的靶细胞-CFSE与未脉冲处理的靶细胞-CFSE。抗原脉冲处理的靶细胞与对照靶细胞之间的频率用作抗原特异性杀伤的读数。
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