High-Throughput Screening for Identification of Novel Innate Immune Activators.
High-Throughput Screening for Identification of Novel Innate Immune Activators.
复制标题
用于鉴定新型先天免疫激活剂的高通量筛选。
DOI：
10.1007/978-1-4939-7237-1_12
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发表时间：
2017
期刊：
影响因子：
--
通讯作者：
DeFilippis,VictorR
中科院分区：
文献类型：
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作者：
Gall,BryanJ;DeFilippis,VictorR
现代药物发现已经采用了体外平台，可以在可处理的时间框架内以可行的成本对大量化合物进行研究。近年来，分子和细胞方法的进步，如基因传递和编辑技术、先进的成像技术、机器人技术和定量分析，极大地帮助了这些努力。因此，对新分子的表型影响的检查可能只受化合物收集的大小的限制。哺乳动物细胞中的先天免疫信号过程特别适合高通量筛选平台，因为它们的激活引起的细胞反应通常导致高水平的转录，可以以生物发光或荧光信号的形式加以利用。此外，靶向激活先天免疫途径是一种适用于多种慢性和急性人类疾病的有价值的治疗策略。在这里，我们描述了一种高通量筛选方法的优化和利用，该方法使用对I型干扰素反应刺激有反应的人类报告细胞。重要的是，所描述的原理和方法可以应用于不同来源的贴壁报告细胞和先天信号通路读数。
Modern drug discovery has embraced in vitro platforms that enable investigation of large numbers of compounds within tractable timeframes and for feasible costs. These endeavors have been greatly aided in recent years by advances in molecular and cell-based methods such as gene delivery and editing technology, advanced imaging, robotics, and quantitative analysis. As such, the examination of phenotypic impacts of novel molecules may only be limited by the size of the compound collection. Innate immune signaling processes in mammalian cells are especially amenable to high-throughput screening platforms since the cellular responses elicited by their activation often result in high level transcription that can be harnessed in the form of bioluminescent or fluorescent signal. In addition, targeted activation of innate immune pathways represents a valuable therapeutic strategy applicable to multiple chronic and acute human diseases. Herein, we describe the optimization and utilization of a high-throughput screening method using human reporter cells reactive to stimulation of the type I interferon response. Importantly, the principles and methods described can be applied to adherent reporter cells of diverse derivation and innate signaling pathway readouts.