Project description:To examine the chromatin accessibility of IELp, single cell epigenomic analysis of IELp from control mice (Cd4 Cre–Lrffl/fl) were determined by scATACseq
Project description:Background: The neonatal (D14-17) murine small intestinal epithelium is enriched for Vγ7+ IEL displaying an ‘immature’ CD122LO cell surface phenotype. We believe these cells are precursors for signature CD122HI Vγ7+ IEL that predominate among small intestinal IEL from postnatal D21. Method: To further characterise this potential precursor-product relationship, CD122hi Vg7+ and CD122lo Vg7+ IEL were purified from individual D14-D17 mice on four independent occasions and compared by total RNAseq. Conclusion: ~3000 genes are differentially expressed between CD122HI and CD122LO murine Vg7+ neonatal IEL
Project description:TCRαβ+CD8αα+ intraepithelial lymphocytes (CD8αα+ αβ IELs), a specialized subset of T cells in the gut epithelium, develop from thymic agonist-selected IEL precursors (IELps). The molecular mechanisms underlying the selection and differentiation of this T cell type in the thymus are largely unknown. Here, we found that Bcl6 deficiency in αβ T cells resulted in nearly the absence of CD8αα+ αβ IELs. BCL6 was expressed by approximately 50% of CD8αα+ αβ IELs but the majority thymic PD1+ IELps post agonist selection; its deficiency blocked early IELp generation in the thymus. Moreover, BCL6 expression in IELps was induced by thymic TCR signaling in an ERK-dependent manner. As a result of Bcl6 deficiency, the precursors of IELps among CD4+CD8+ double positive (DP) thymocytes exhibited increased apoptosis during agonist selection, and impaired IELp differentiation and maturation. Taken together, our results elucidate BCL6 as a crucial transcription factor during the thymic development of CD8αα+ αβ IELs.
Project description:To examine how the cluster composition of CD8aa IEL and their transcriptomic signatures were affected by LRF disruption, single-cell gene expression of CD8aa IEL from control (Cd4 Cre–Lrffl/fl) and CD8aa splenocytes from LRF KO (Cd4 Cre+Lrffl/fl ) mice were determined by scRNAseq.
Project description:Background: Recently have found that Btnl1 expressed by murine enterocytes shapes the local TCR-Vγ7+ compartment by driving their clonotypic expansion and phenotypic maturation from CD122LO to CD122HI Vγ7+ cells. The neonatal (D14-17) murine small intestinal epithelium is enriched for Vγ7+ IEL displaying an â??immatureâ?? CD122LO cell surface phenotype. We believe these cells are precursors for CD122HI Vγ7+ IEL. Method: To further characterise this putative product-progenitor relationship, CD122hi Vg7+ and CD122lo Vg7+ IEL were purified from individual D14-D17 mice on four independent occasions and compared by total RNAseq. Conclusion: >3000 genes are differentially expressed between CD122HI and CD122LO murine Vg7+ neonatal IEL 4 independent Paired samples of CD122hi Vg7+ and CD122lo Vg7+ IEL were purified from the small intestinal epithelium of pooled 14-17 day old mice. RNA was isolated from sorted samples. Gene expression analysis was performed by total RNAseq.