Perez Omar D, Nolan Garry P
The Baxter Laboratory for Genetic Pharmacology, Department of Microbiology and Immunology, Stanford University, School of Medicine, Stanford, CA 94305, USA.
Immunol Rev. 2006 Apr;210:208-28. doi: 10.1111/j.0105-2896.2006.00364.x.
Understanding a molecular basis for cellular function is a common goal of biomedicine. The complex and dynamic cellular processes underlying physiological processes become subtly or grossly perturbed in human disease. A primary objective is to demystify this complexity by creating and establishing relevant model systems to study important aspects of human disease. Although significant technological advancements over the last decade in both genomic and proteomic arenas have enabled progress, accessing the complexity of cellular interactions that occur in vivo has been a difficult arena in which to make progress. Moreover, there are extensive challenges in translating research tools to clinical applications. Flow cytometry, over the course of the last 40 years, has revolutionized the field of immunology, in both the basic science and clinical settings, as well as having been instrumental to new and exciting areas of discovery such as stem cell biology. Multiparameter machinery and systems exist now to access the heterogeneity of cellular subsets and enable phenotypic characterization and functional assays to be performed on material from both animal models and humans. This review focuses primarily on the development and application of using activation-state readouts of intracellular activity for phospho-epitopes. We present recent work on how a flow cytometric platform is used to obtain mechanistic insight into cellular processes as well as highlight the clinical applications that our laboratory has explored. Furthermore, this review discusses the challenges faced with processing high-content multidimensional and multivariate data sets. Flow cytometry, as a platform that is well situated in both the research and clinical settings, can contribute to drug discovery as well as having utility for both biomarker and patient-stratification.
理解细胞功能的分子基础是生物医学的一个共同目标。生理过程背后复杂而动态的细胞过程在人类疾病中会受到轻微或严重的干扰。一个主要目标是通过创建和建立相关模型系统来研究人类疾病的重要方面，从而揭开这种复杂性的神秘面纱。尽管在过去十年中基因组学和蛋白质组学领域取得了重大技术进步并推动了研究进展，但了解体内发生的细胞相互作用的复杂性一直是一个难以取得进展的领域。此外，将研究工具转化为临床应用面临诸多挑战。在过去40年里，流式细胞术在基础科学和临床环境中都彻底改变了免疫学领域，并且对诸如干细胞生物学等新的令人兴奋的发现领域也起到了重要作用。现在存在多参数仪器和系统来分析细胞亚群的异质性，并能够对来自动物模型和人类的材料进行表型特征分析和功能测定。本综述主要关注利用细胞内活性的磷酸化表位激活状态读数的开发和应用。我们展示了最近关于如何使用流式细胞术平台来深入了解细胞过程的机制的研究工作，并突出了我们实验室所探索的临床应用。此外，本综述讨论了处理高含量多维和多变量数据集所面临的挑战。流式细胞术作为一个在研究和临床环境中都有良好定位的平台，可有助于药物发现，并且对生物标志物和患者分层都有用处。
