Supplementary MaterialsSupplementary Information 41467_2020_15413_MOESM1_ESM. transcription factors regulate the balance between effector and memory cell differentiation during T cell activation. Here, we analyse the role of the TRAF2-/NCK-interacting kinase Deferitrin (GT-56-252) (TNIK), a signaling molecule downstream of the tumor necrosis factor superfamily receptors such as CD27, in the regulation of CD8+ T cell fate during acute infection with lymphocytic choriomeningitis virus. Priming of CD8+ T cells induces a TNIK-dependent nuclear translocation of -catenin with consecutive Deferitrin (GT-56-252) Wnt pathway activation. TNIK-deficiency during T cell activation results in enhanced differentiation towards effector cells, glycolysis and apoptosis. TNIK signaling enriches for memory precursors by favouring symmetric over asymmetric cell division. This enlarges the pool of memory CD8+ T cells and increases their capacity to expand after re-infection in serial re-transplantation experiments. These findings reveal that TNIK is an important regulator of effector and memory T cell differentiation and induces a population of stem cell-like memory T cells. (test, nonsignificant compared to before priming impairs CD8+ T-cell memory formation.a Gp33-Tet+ CD8+ T-cell frequency in blood of 200 pfu LCMV-WE-infected test, nonsignificant deletion (Supplementary Fig.?1a). Purified splenic test, nonsignificant test, nonsignificant and involved in differentiation34 and involved in asymmetric cell division35,36 were expressed at higher levels in KO p14 T cells (Fig.?4d; Supplementary Fig.?5h). Transcriptional regulators determining T-cell development and function such as and and that are involved in the Wnt pathway and (CD107), were expressed at higher levels in WT memory p14 memory T cells. In contrast, the transcription factor regulating effector fate41, were expressed at significantly lower levels in KO vs WT p14 T cells (Fig.?6a; Supplementary Fig.?7a). gene expression was significantly higher in?AdTf WT vs KO p14 T cells 48?h p.i., confirming our in vitro data (Supplementary Fig.?7b). However, Wnt target genes were not differentially expressed in the NGS analysis of KO vs WT p14 T cells day 6 p.i., suggesting that Wnt target genes may be induced very early after T-cell stimulation. expression and the expression of genes associated with T-cell effector function (test, nonsignificant test, nonsignificant and other molecules associated with differentiation to effector cells such as and are upregulated in TNIK KO effector p14 T cells. Notch and Wnt pathways are highly conserved interrelated signaling pathways that reciprocally control cell fate57. In CD8+ T cells, Notch signaling promotes effector differentiation while inhibiting the signaling pathways promoting memory T-cell formation6. Moreover, Notch activates the PI3K/Akt/mTOR pathway that is critical for metabolic conversion to glycolysis, allowing rapid proliferation and acquisition of effector function by T cells47. Importantly, GSE analysis of TNIK-deficient effector cells revealed a significantly higher expression of genes involved in the PI3K/Akt pathway, suggesting that Akt and mTOR kinases contribute to the increased glycolysis. Wnt signaling favors the differentiation into memory precursor cells10. The Wnt target genes and are preferentially expressed in TN and in TCM, but not in TEFF cells58. Moreover, activation of the Wnt pathway in vitro suppressed the antigen-induced expression Eomes and inhibited differentiation to effector T cells. This arrested differentiation favored the generation of TCM and T memory stem cells that are characterized by a high proliferative capacity upon TCR re-stimulation53,59. Further, allele or littermate controls were generated. Genotyping primers (Supplementary Table?1) were designed by KOMP Repository (Design ID: 49289). Per oral Deferitrin (GT-56-252) (p.o.) administration of tamoxifen (200?mg?kg?1 day?1) on 5 consecutive days allowed Cre-mediated TNIK deletion. By crossing were generated. P14 TCR mice were crossed with mice and littermate controls were Rabbit polyclonal to RAB1A infected with 200 plaque-forming units (pfu) LCMV-WE. Alternatively, 1??105 MACS-purified p14 CD8+ T cells from p14;test (one-tailed, two-tailed). Significant differences in KaplanCMeier survival curves were determined using the log-rank test (two-tailed). Data are represented as means??standard error of the mean (SEM) as indicated in the legend. thanks the anonymous reviewer(s) for their contribution to the peer review of this work. Peer reviewer reports are available. Publishers note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations. These authors contributed equally: Carla A. Jaeger-Ruckstuhl, Magdalena Hinterbrandner. Supplementary information Supplementary information is available for this paper at 10.1038/s41467-020-15413-7..