Department of Medicine/Nephrology, Stanford University, Palo Alto, California.
Institute for Immunity, Transplantation and Infection, Stanford University, Stanford, California.
Am J Transplant. 2019 May;19(5):1278-1287. doi: 10.1111/ajt.15316. Epub 2019 Mar 20.
Recently developed single-cell profiling technologies hold promise to provide new insights including analysis of population heterogeneity and linkage of antigen receptors with gene expression. These technologies produce complex data sets that require knowledge of bioinformatics for appropriate analysis. In this minireview, we discuss several single-cell immune profiling technologies for gene and protein expression, including cytometry by time-of-flight, RNA sequencing, and antigen receptor sequencing, as well as key considerations for analysis that apply to each. Because of the critical importance of data analysis for high parameter single cell analysis, we discuss essential factors in analysis of these data, including quality control, quantification, examples of methods for high dimensional analysis, immune repertoire analysis, and preparation of analysis pipelines. We provide examples of, and suggestions for, application of these innovative methods to transplantation research.
最近开发的单细胞分析技术有望提供新的见解,包括对群体异质性的分析以及抗原受体与基因表达的关联。这些技术产生了需要生物信息学知识才能进行适当分析的复杂数据集。在这篇小综述中,我们讨论了几种单细胞免疫分析技术,用于基因和蛋白质表达,包括飞行时间流式细胞术、RNA 测序和抗原受体测序,以及适用于每种技术的分析的关键考虑因素。由于高参数单细胞分析的数据分析至关重要,我们讨论了分析这些数据的基本因素,包括质量控制、定量、高维分析方法的示例、免疫受体库分析以及分析管道的准备。我们提供了将这些创新方法应用于移植研究的示例和建议。