Xpressed in each tissues (Fig. 2B, C). The expression differences in
Xpressed in each tissues (Fig. 2B, C). The expression differences in these key genes have been comparable among 3 donors (Fig. 2C). The genes differentially expressed by human CD69+ and CD69-TEM cells (Fig. 2C) integrated crucial molecules linked with mouse CD8+TRM from infection models (MackayCell Rep. Author manuscript; readily available in PMC 2017 October 18.Author Manuscript Author Manuscript Author Manuscript Author ManuscriptKumar et al.Pageet al., 2016; Mackay et al., 2013; Skon et al., 2013; Wakim et al., 2012). Notably, downregulation of S1PR1 and its related transcription element KLF2 are required for CD8+TRM establishment in mice (Skon et al., 2013), and we found striking downregulation of S1PR1 (8-16-fold) and KLF2 (2-16-fold) transcripts for all CD69+ compared with CD69- subsets in every single donor for each CD4+ and CD8+T cells in lung and spleen (Fig. 2D). In addition, human CD8+CD69+ subsets exhibited upregulation of ITGAE (CD103), ITGA1 (CD49a), ICOS, as well as the transcription issue IRF4, also identified to be upregulated by mouse CD8+TRM in distinct systems (Mackay and Kallies, 2017). Collectively, these outcomes show that the CD69+ tissue memory T cells comprise a transcriptionally distinct subset enriched for characteristics of tissue residency. We further compared the transcriptional profiles of tissue memory T cell subsets with circulating TEM cells isolated in the blood of 3 healthier volunteers. PCA evaluation applying the gene signature in Fig. 2C resulted in clustering of blood TEM with CD69- tissue TEM, distinct from CD69+ samples which clustered collectively (Fig. 2E). By contrast, PCA evaluation using an equal quantity of randomly chosen genes as a adverse handle yielded no clustering pattern (Fig. S1). This grouping suggests that CD69 expression by memory T cells in tissues distinguishes circulating memory subsets from those retained in tissues. A core gene signature of human CD69+ memory T cells Depending on the gene expression analysis above, we identified 31 core genes with constant significant differential expression by CD4+ and CD8+ CD69+ compared with all the corresponding CD69- subset from lung, spleen, and blood (Fig. 3A and Table S2). This core signature incorporated upregulation of the adhesion markers ITGAE (CD103) and ITGA1 (CD49a), the chemokine-receptors CXCR6 and CX3CR1, and molecules with known inhibitory functions in T cells including PDCD1 (PD-1) (Barber et al., 2006), the dualspecificity phosphatase DUSP6 that turns off MAP Kinase signaling (MCP-1/CCL2, Mouse (HEK293) Bertin et al., 2015), and IL10 (IL-10). Downregulated genes inside the core signature integrated S1PR1 and its connected transcription element KLF2, which collectively manage T cell homing and tissue retention (Skon et al., 2013), the associated Kruppel-like transcription element KLF3, the lymph node homing UBE2M Protein Storage & Stability receptor SELL (CD62L), at the same time as RAP1GAP1 and RGS1, G protein signaling genes that modulate T cell trafficking (Gibbons et al., 2011). Pathways represented inside the core signature contain these controlling T cell adhesion and migration, proliferation, improvement, and activation (Table S3) that interconnect as diagrammed in Fig. 3B. A lot of on the upregulated genes map downstream of TCR signaling, like CD69, adhesion molecules (ITGA1, ITGAE, CRTAM), and activation-induced molecules IL2 (IL-2), IL10 (IL-10), and PDCD1 (PD-1) that could regulate proliferation (Fig. 3B). Differential upregulation or downregulation of specific chemokines and chemokine receptors (CXCL13, CXCR6, CX3CR1, SELL, S1PR1) and modu.
