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:Mouse small intestine intraepithelial lymphocytes (IEL) that express a ab TCR and CD8aa homodimers are an enigmatic T cell subset, as their specificity and in vivo function remain to be defined. To gain insight into the nature of these cells, we performed global gene expression profiling using microarray analysis, combined with PCR and flow cytometry to determine the level of expression of selected genes. Using these methods, TCR ab+ CD8aa IEL were compared to their TCR ab+ CD8b+ and TCR gd+ counterparts. Keywords: Cell type comparison
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:We report the application of RNA-Seq technology for high-throughput gene expression profiling of intestinal intraepithelial lymphocytes (IEL) that were incubated for 24 hrs in the presence or absence of recombinant osteopontin.
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