SMART-seq of PD1+ and PD1- CD8 T cells derived from Setx L389S (Setx KI) mice

The goal is to understand the nature of PD-1+ CD8 T cells that are detected in the peripheral blood of ALS4 mice (expressing the mutation SetxL389S, also called Setx KI mice). We performed a SMART (Switching Mechanism At the end of the 5'-end of the RNA Transcript)-sequencing analysis of the transcriptome of effector memory CD8 T cells expressing or not PD-1 from the blood of symptomatic KI animals. Amongst the transcripts differentially expressed between these two populations, we detected upregulation of Tox (Thymocyte Selection Associated High Mobility Group Box) in PD-1+ as compared to PD-1- CD8 T cells. 500 CD44+CD62L-PD1+ and 500 CD44+CD62L-PD-1-CD8 T cells from 10-12 month old KI mice were sorted using a BD FACSAria with 3 or 5 lasers (BD Bioscience). Three individual mice were analyzed.本研究旨在明确ALS4小鼠——即携带SetxL389S突变的Setx基因敲入（KI）小鼠——外周血中检测到的PD-1阳性CD8 T细胞（PD-1+ CD8 T cells）的生物学特性。我们对症状发作期KI小鼠血液中表达PD-1与不表达PD-1的效应记忆CD8 T细胞的转录组，开展了SMART测序（RNA转录本5'末端转换机制测序，Switching Mechanism At 5' End of RNA Transcript Sequencing）分析。在这两类细胞群的差异表达转录本中，我们发现相较于PD-1阴性CD8 T细胞，PD-1阳性CD8 T细胞中Tox（胸腺细胞选择相关高迁移率族盒蛋白）的表达水平显著上调。本研究从10至12月龄的KI小鼠体内，分选得到500个CD44阳性、CD62L阴性、PD-1阳性的CD8 T细胞，以及500个CD44阳性、CD62L阴性、PD-1阴性的CD8 T细胞；分选操作采用配备3或5个激光器的BD FACSAria流式细胞分选仪（BD Bioscience）完成，共对3只独立的实验小鼠进行了上述分析。