一种在原位同时可视化病毒特异性CD8 + T细胞和病毒感染细胞的技术。
A technique to simultaneously visualize virus-specific CD8+ T cells and virus-infected cells in situ.
作者信息
Li Qingsheng, Skinner Pamela J, Duan Lijie, Haase Ashley T
机构信息
Department of Microbiology, Medical School, University of Minnesota, USA.
出版信息
J Vis Exp. 2009 Aug 13(30):1561. doi: 10.3791/1561.
Abstract
The numbers and locations of virus-specific CD8+ T cells relative to the numbers and locations of their infected cell targets is thought to be critical in determining outcomes that range from clearance to chronic persistent infections. We describe here a method for assessing the spatial and quantitative relationships between immune effector (E) virus-specific CD8+ T cells and infected targets (T) that combines in situ tetramer (IST) staining to detect virus-specific CD8+ T cells and in situ hybridization (ISH) to detect viral-RNA+ cells in the tissues where the battle between immune defenses and infection takes place. The combination of IST staining and ISH, abbreviated ISTH, enables visualization and mapping of the locations of immune effector cells and targets, and facile determination of E:T ratios. These parameters in turn can then be used to determine the relationships between spatial proximity, and the timing and magnitude of the immune response that predict outcomes in early infection.
摘要
相对于其被感染细胞靶标的数量和位置，病毒特异性CD8 + T细胞的数量和位置被认为在决定从清除感染到慢性持续性感染等一系列结果中起着关键作用。我们在此描述一种评估免疫效应细胞（E）病毒特异性CD8 + T细胞与被感染靶标（T）之间空间和定量关系的方法，该方法结合了原位四聚体（IST）染色以检测病毒特异性CD8 + T细胞，以及原位杂交（ISH）以检测免疫防御与感染发生战斗的组织中的病毒RNA +细胞。IST染色和ISH的结合（缩写为ISTH）能够可视化并绘制免疫效应细胞和靶标的位置，并轻松确定E:T比率。这些参数进而可用于确定空间接近度、免疫反应的时间和强度之间的关系，这些关系可预测早期感染的结果。
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