Transcriptional Changes during T cell development following conditional loss of Bcl6

In this study we investigated the changes during T cell development in Rag knockout Bcl6 conditional knockout E16.5 thymi compared to Control E16.5 thymi. We measured the transcription changes by RNA seq in three different populations including CD25+, CD8+CD4- (ISP) and CD8+CD4+ (DP) thymocytes. Embryonic day (E16.5) thymi from Rag knockout Bcl6 conditional knockout (Ragfl/flBcl6LckCre) embryos were used for this study. CD25 positive thymocytes were isolated from Control (RagKOBclfl/flLckCre-) and RagKOBcl6coKO (RagKOBcl6fl/flLckCre+). Some RagKOBcl6coKO thymi were treated with anti-CD3 for 2days to promote differentiation following a TCR mimicking stimulus. CD8+CD4- (ISP) cells and CD8+CD4+ (DP) cells were isolated from these treated thymi. RNA-Seq was carried out on the CD25 positive, ISP and DP populations from Control and RagKOBcl6coKO samples.

本研究以Rag基因敲除Bcl6条件性敲除胚胎的E16.5胸腺与对照胚胎的E16.5胸腺为研究对象，探究二者T细胞发育过程中的转录变化。我们通过RNA测序（RNA-seq）检测了三类胸腺细胞群的转录变化，分别为CD25阳性胸腺细胞、CD8阳性CD4阴性（ISP）细胞以及CD8阳性CD4阳性（DP）细胞。本研究使用的实验样本来自Rag基因敲除Bcl6条件性敲除（Ragfl/flBcl6LckCre）胚胎的E16.5胸腺。研究人员从对照胚胎（RagKOBclfl/flLckCre-）与RagKOBcl6条件性敲除（RagKOBcl6fl/flLckCre+）胚胎中分离得到CD25阳性胸腺细胞。部分RagKOBcl6条件性敲除胸腺经抗CD3抗体处理2天，以模拟T细胞受体（TCR）刺激并促进细胞分化，随后从该处理组胸腺中分离得到CD8阳性CD4阴性（ISP）细胞与CD8阳性CD4阳性（DP）细胞。最终，研究人员对对照样本与RagKOBcl6条件性敲除样本中的CD25阳性、ISP及DP细胞群开展了RNA测序分析。