Institute for Immunity, Transplantation, and Infection Operations, USA.
Curr Opin Endocrinol Diabetes Obes. 2010 Aug;17(4):322-8. doi: 10.1097/MED.0b013e32833ada91.
This article will review new technologies used to characterize the immune phenotype of cells and serum for potential use in studies of autoimmunity.
One area of recent development in studies of immune phenotyping is the contrast between cells of the immune system at rest and following activation. This simply involves comparing these cells at rest and following ligand-induced activation and measuring signaling system activation (phosphoepitope identification) or intracellular cytokine production or activation-induced gene expression. Preliminary data using these techniques have begun to identify signatures of disease (biomarkers) that are only seen when using these activation-induced assays. One of the most exciting new technologies, cytometry by time-of-flight mass spectrometry, couples a flow cytometer to a mass spectrometer, allowing many more parameters to be analyzed per cell, and without spillover between assay reagents, compared to conventional optical flow cytometry (heavy ions, mass, replaces fluorophore readout). Another new technology to analyze soluble proteins, bead-based immunoassays, can simultaneously measure up to 75 soluble analytes in a multiplexed array. Other technologies provide similar innovations in terms of analytical content, throughput, and miniaturization.
We believe that new cellular genomic and protein-based technologies can provide key insights into autoimmune disease pathogenesis, progression, and therapy, and that these assays need to be applied in a systematic way to samples from patients with autoimmune diseases.
本文将回顾用于描述细胞和血清免疫表型的新技术,以期将其应用于自身免疫研究。
免疫表型研究的一个新领域是比较免疫系统细胞在静止和激活后的状态。这只需比较这些细胞在静止和配体诱导激活后的状态,并测量信号系统激活(磷酸化表位鉴定)或细胞内细胞因子产生或激活诱导的基因表达。使用这些技术的初步数据已开始确定仅在使用这些激活诱导测定时才出现的疾病特征(生物标志物)。最令人兴奋的新技术之一是飞行时间质谱流式细胞术,它将流式细胞仪与质谱仪相结合,与传统的光学流式细胞术相比,每个细胞可分析更多的参数,且测定试剂之间没有串扰(重离子、质量,取代荧光团读取)。另一种用于分析可溶性蛋白的新技术是基于珠的免疫测定法,可在一个多重阵列中同时测量多达 75 种可溶性分析物。其他技术在分析内容、通量和小型化方面提供了类似的创新。
我们认为,新的细胞基因组和基于蛋白质的技术可以为自身免疫性疾病的发病机制、进展和治疗提供重要的见解,并且这些检测需要以系统的方式应用于患有自身免疫性疾病的患者的样本中。