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: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.
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:Aire is a transcriptional regulator that induces promiscuous expression of thousands of tissue-restricted antigen (TRA) genes in medullary thymic epithelial cells (mTECs). While the target genes of Aire are well characterized, the transcriptional programs regulating its own expression remain elusive. We used Affymetrix microarrays to analyze the gene expression patterns of Aire expressing cells (mature mTECs and Thymic B cells) and compared them to control counterparts, namely immature mTECs, cortical Thymic epithelial cells and splenic B cells of tissue-restricted antigen (TRA) genes in medullary thymic epithelial cells (mTECs). While the target genes of Aire are well characterized, the transcriptional programs regulating its own expression remain elusive. We’ve used Assay for transposase-accessible chromatin using sequencing (ATAC-Seq) on the different thymic epithelial cell populations to assess chromatin accessibility around the Aire locus in these cells. Moreover, we’ve used the indexing-first chromatin immunoprecipitation (iChIP) technique to assess the occupancy of the Irf8 transcription factor in the Aire locus