在这篇文章里,Roederer和他的团队提出了一种在BioMark平台使用多重TaqMan试剂对单个T细胞进行分析的方法。首先,使用BD FACSAria™,根据细胞的标记将其分离以得到单细胞。接下来,Roederer细化其方法学,确保获得足量的RNA以进行单细胞的表达谱分析。Roederer等展示了低至一个mRNA转录本的检测极限(limit of detection, LOD),结果表明BioMark的高灵敏度和宽的动态范围,可以检测单细胞当量的mRNA。
该团队在单细胞水平监测了T细胞的激活,发现了群体中CD4+细胞的亚群。实际上,CD4+细胞表达类似CSCR5/CCL5和DPPR/TIA1这样的基因是稀有事件,并非之前使用大量mRNA观察到的是普遍事件
通过比较CD154+和CD154- T细胞对葡萄球菌肠毒素B(SEB, Staphlyococcal enterotoxin B)的反应,(细胞)的异质性被进一步确立。当被刺激时,分析显示有一组基因和T细胞的激活强烈相关,但是,只有5-20%的T细胞拥有可以结合SEB的受体Vβ链变体。
在这项研究中,细胞异质性被证明有很强的免疫相关性。Roederer和他的团队正继续推进系统的、单细胞的方法以更好地理解中枢免疫系统中细胞和分子的相互作用。
参考文献
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