| 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 ...
[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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| Quantification of T Cell Antigen-specific Memory Responses in Rhesus Macaques, Using Cytokine Flow Cytometry (CFC, also Known as ICS and ICCS): from Assay Set-up to Data Acquisition
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Author:
Date:
2014-04-20
[Abstract] What was initially termed ‘CFC’ (Cytokine Flow Cytometry’) is now more commonly known as ‘ICS’ (Intra Cellular Staining), or less commonly as ‘ICCS’ (Intra Cellular Cytokine Staining). The key innovations were use of an effective permeant (allowing intracellular staining), and a reagent to disrupt secretion (trapping cytokines, thereby enabling accumulation of detectable intracellular signal). Because not all researchers who use the technique are interested in cytokines, the ‘ICS’ term has gained favor, though ‘CFC’ will be used here.
CFC is a test of cell function, exposing ...
[摘要] 最初称为"CFC"（细胞因子流式细胞术）的现在更通常称为"ICS"（细胞内染色），或更不常称为"ICCS"（细胞内细胞因子染色）。关键的创新是使用有效的渗透物（允许细胞内染色）和试剂破坏分泌（捕获细胞因子，从而能够累积可检测的细胞内信号）。因为不是所有使用该技术的研究人员都对细胞因子感兴趣，'ICS'术语已经受到青睐，尽管这里将使用"CFC"。
CFC是细胞功能的测试，将淋巴细胞暴露于培养物中的抗原，然后测量任何诱导的细胞因子应答。处理测试培养物以用用荧光标记物标记的单克隆抗体染色细胞，并使用多聚甲醛化学固定细胞和净化样品。
CFC提供流式细胞术的功能，包括批量采样和多参数互相关，用于抗原特异性记忆反应的分析。使用CFC的研究者能够表型表征用测试抗原和表型亚型（例如，CD4 + 或CD8 + T细胞）培养的细胞，确定高于背景水平的产生频率的细胞因子。
与ELISPOT和Luminex方法相反，CFC可以关联来自特定的，表型 - 表征的细胞的多种细胞因子的产生。 ...
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