Project description:To examine how the transcriptome of circulating memory CD8 T cells isolated from the spleen are reprogrammed in the lung airways, influenza nucleoprotein-specific CD8 T cells from the spleen were FACS sorted and RNA isolated for RNA-seq or the cells were transferred to the lung airways and allowed to rest for 2 days. Following 2 days, donor influenza nucleoprotein-specific CD8 T cells were FACS sorted and RNA isolated for RNA-seq..
Project description:To examine differences in chromatin accessibility of resident memory CD8 T cells in the lung, influenza nucleoprotein-specific CD8 T cells from the airways , lung parenchyma, and spleen were FACS sorted and DNA isolated for ATAC-seq.
Project description:To examine differences in the transcriptome of resident memory CD8 T cells in the lung, influenza nucleoprotein-specific CD8 T cells from the airways , lung parenchyma, and spleen were FACS sorted and RNA isolated for RNA-seq.
Project description:To examine differences in the transcriptome of resident memory CD8 T cells in the lung, influenza nucleoprotein-specific CD8 T cells from the airways , lung interstitium, lung vasculature, and spleen were FACS sorted and RNA isolated for RNA-seq. In some experiments, FACS sorted cells were transferred into the airways, peritoneum, or in vitro culture before subsequent isolation and RNA isolation.
Project description:To examine differences in the transcriptome of resident memory CD8 T cells in the lung, influenza nucleoprotein-specific CD8 T cells from the airways , lung interstitium, and spleen were FACS sorted and RNA isolated for RNA-seq.
Project description:To examine differences in chromatin accessibility of resident memory CD8 T cells in the lung, influenza nucleoprotein-specific CD8 T cells from the airways , lung interstitium, and spleen were FACS sorted and ATAC-seq performed.
Project description:Tissue resident memory (Trm) represent a newly described memory T cell population. We have previously characterized a population of Trm that persists within the brain following acute virus infection. Although capable of providing marked protection against a subsequent local challenge, brain Trm do not undergo recall expansion following dissociation from the tissue. Furthermore, these Trm do not depend on the same survival factors as the circulating memory T cell pool as assessed either in vivo or in vitro. To gain greater insight into this population of cells we compared the gene-expression profiles of Trm isolated from the brain to circulating memory T cells isolated from the spleen following an acute virus infection. Trm displayed altered expression of genes involved in chemotaxis, expressed a distinct set of transcription factors and overexpressed several inhibitory receptors. Cumulatively, these data indicates that Trm are a distinct memory T cell population disconnected from the circulating memory T cell pool and displaying a unique molecular signature which likely results in optimal survival and function within their local environment. 13 samples were analyzed: 5 replicates of memory OT-I CD8+.CD103- T cells isolated from the spleen of mice on day 20 p.i. with VSV-OVA. 5 replicates of memory OT-I CD8+CD103+ T cells isolated from the brain of mice on day 20 p.i. with VSV-OVA; and 3 replicates of memory OT-I.CD8+ CD103- T cells isolated from the brain of mice on day 20 p.i. with VSV-OVA
Project description:We generated a single-cell transcriptomic resource dataset encompassing the gene-expression patterns of circulating and small intestine intraepithelial CD8+ T cells in response to viral infection. Our analyses revealed a core transcriptional program shared between circulating memory and tissue-resident memory (TRM) cells, along with key differences in the kinetics and magnitude of gene expression between these two memory CD8+ T lymphocyte subtypes. Moreover, we elucidated previously unappreciated heterogeneity within the small intestine intraepithelial CD8+ T cell pool at multiple time points following infection.
Project description:Memory CD8+ T cells are indispensable for maintaining long-term immunity against intracellular pathogens and tumors. Despite their presence in oxygen-deprived tissues of infection sites or tumors, the impact of local oxygen pressure on memory CD8+ T cells has remained largely unclear. We sought to elucidate how oxygen pressure impacted memory CD8+ T cells arising after infection with Listeria monocytogenes-OVA. Our data revealed that reduced oxygen pressure during in vitro culture switched CD8+ T cell metabolism from an OXPHOS to a glycolytic phenotype. Quantitative proteomic analysis showed that limiting oxygen conditions increased the expression of glucose transporters and components of the glycolytic pathway, while decreasing TCA cycle and mitochondrial respiratory chain proteins. The altered CD8+ T cell metabolism did not affect the expansion potential, but enhanced the granzyme B and IFN- production capacity. Memory CD8+ T cells cultured under low oxygen pressure were able to persist long-term in vivo and provided protection against bacterial rechallenge. Taken together, our study indicates that strategies of cellular immune therapy may benefit from reducing oxygen during culture to develop memory CD8+ T cells with superior effector functions .
Project description:The interleukin-23 (IL-23) pathway plays a critical role in the pathogenesis of multiple chronic inflammatory disorders, however, inter-individual variability in IL-23-induced signal transduction in circulating human lymphocytes has not been well-defined. In this study, we observed marked, reproducible inter-individual differences in IL-23 responsiveness (measured by STAT3 phosphorylation) in peripheral blood CD8+CD45RO+ memory T and CD3+CD56+ NKT cells. To define mechanisms that might be contributing to the differential IL-23-induced STAT3 activation between individuals, we examined mRNA expression differences in CD8+CD45RO+ memory T cells between IL-23 responsive and non-responsive individuals. We analyzed unstimulated and IL-23 stimulated FACS sorted CD8+CD45RO+ memory T cells from two individuals demonstrating robust IL-23 responsiveness, and two individuals demonstrating low IL-23 responsiveness, using the Affymetrix Human Exon 1.0 ST platform. Array data was processed by Affymetrix Expression Console software. No techinical replicates were performed.