| Murine in vitro Memory T Cell Differentiation
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Author:
Date:
2014-07-05
[Abstract] Upon pathogen encounter, naïve CD8+ T cells are primed and undergo massive clonal expansion. A fraction of effector CD8+ T cells remains during the contraction phase and differentiate into memory T cells critical for mounting robust recall responses in response to secondary infection. Low frequency of memory T cells in vivo is a major obstacle to investigate their functional aspects including migration capacity and genetic regulation. Here, we describe detailed protocol for memory T cell differentiation developed by von Andrian’s group to generate large number of CD44hiCD62Lhi antigen-specific memory T cells in vitro.
[摘要] 在病原体遭遇时，初始CD8 + T细胞被引发并经历大量克隆扩增。 效应CD8 + sup T细胞的一部分在收缩阶段期间保持并分化为响应于继发感染而对于安装鲁棒回忆反应至关重要的记忆T细胞。 体内低频记忆T细胞是研究其功能方面（包括迁移能力和遗传调节）的主要障碍。 在这里，我们描述由von Andrian的小组开发的记忆性T细胞分化的详细方案，以在体外产生大量的CD44 - 高 - 抗原特异性记忆T细胞 。
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| Murine in vivo CD8+ T Cell Killing Assay
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Author:
Date:
2014-07-05
[Abstract] Antigen-specific killing ability of effector CD8+ T cells is critical for protective immunity against infection. Here, we describe in vivo cytotoxic T cell assay to examine effector function of antigen-specific CD8+ T cells. Mice infected with Listeria monocytogenes (L. monocytogenes) expressing chicken ovalbumin as a model antigen mount ovalbumin-specific CD8+ T cell responses. Effector CD8+ T cell function in vivo 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-CFSElow vs. unpulsed target cells-CFSEhi. ...
[摘要] 效应CD8 + T细胞的抗原特异性杀伤能力对于感染的保护性免疫是关键的。 在这里，我们描述了在体内细胞毒性T细胞测定来检查抗原特异性CD8 + T细胞的效应子功能。 用表达鸡卵白蛋白作为模型抗原的单核细胞增生性李斯特菌（单核细胞增生李斯特氏菌）感染的小鼠装载卵清蛋白特异性CD8 + T细胞应答。 通过将OVA肽脉冲的靶细胞与对照靶细胞混合转移到先前免疫的小鼠中来测定体内效应物CD8 + T细胞功能。 CFSE表达水平的差异清楚地标记两种不同的群体：抗原脉冲的靶细胞-CFSE 低与未突发的靶细胞-FSE hi 。 抗原脉冲的靶细胞和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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