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Gifts on Sale. Site Search. In contrast, a significantly higher proportion of Bach2 -transduced 2C T cells expressed CD62L and CD27, but a significantly lower fraction expressed Klrg1, in the DLN and spleen compared with non-transduced 2C cells in the same organs of the same recipients. Representative data from six mice in three independent experiments are shown. Representative data from 9—12 mice in three independent experiments are shown. We also examined the effect of Bach2 knockdown on memory T-cell development.

The approach was the same as outlined as in Fig. To further explore the mechanism underlying the observed effect of Bach2 on memory T-cell development and response, we examined whether Bach2 affects T-cell proliferation.

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In the IL-7 culture, the proportion of transduced versus non-transduced 2C cells remained stable regardless of whether the 2C T cells were transduced with vector or Bach2 Fig. Similarly, the proportion of vector-transduced versus non-transduced 2C cells remained stable in the IL-2 cultures. When the cells were lableled with eFluor and followed over time, Bach2 -transduced 2C T cells diluted the flourescent dye more extensively than non-transduced 2C T cells Fig. Shown are representative histograms of eFluor from one of the two experiments. The levels of the indicated TFs were assayed by western blotting.

Shown are representative western blotting and the average expression level quantified from three independent experiments. DNA was used to amplify different parts of the Prdm1 e and Id3 f promoter region indicated by i—v. The location of the predicted Bach2 binding motif was indicted as triangle.

Data are from two independent experiments, error bar: s. We verified this regulatory motif by showing that overexpression of Bach2 suppressed expression of Prdm1 but stimulated expression of Id3 at transcriptional Table 3 and translational levels Fig. Further supporting the regulatory motif, overexpression of Prdm1 significantly reduced the Bach2 -mediated proliferation of activated T cells Supplementary Fig. As Prdm1 is known to inhibit T-cell proliferation whereas Id3 stimulates survival of effector T cells 15 , 16 , we determined the effect of Bach2 overexpression on molecules that regulate cell cycle and survival.

These results suggest that Bach2 regulates memory T-cell development and recall proliferation by regulating cell cycle control possibly through Prdm1 and Id3. Our approach is valid based on the following considerations. Conversely, knockdown of Sox4 and Bach2 inhibited the recall proliferation of the transduced memory T cells Fig. Overexpression of Bhlhe40 and Runx2 inhibited the in vivo recall proliferation, although no significant change was observed in in vitro recall response and in knockdown assay Fig. Third, compared with the traditional method of differential gene expression analysis 2 , 3 , which generates a long list of candidates using fold-change-based approaches, our network methods identify and rank order TFs according to their statistical importance.

Reduction of hundreds of TFs to two-dozen key TFs makes direct experimental validation more manageable. The network approach and methodologies developed here can be applied to any phenotypic transition, such as effector T cells and exhausted T cells, to identify novel transcriptional modules and TFs.

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Our network and perturbation studies have now greatly expanded the understanding of the mode of interactions to the top 21 TFs. Our analysis reveals a dense overlapping regulation among the key TFs Fig. This mode of regulation is essential for sensing multiple external signals and integrate them into distinct cell fate outcomes Our analysis also shows complex regulations with both feedback and feed-forward motifs among the key TFs Table 3 and Fig. Although we do not know the precise causes underlying the observed opposite effects, the following differences between the two studies may provide part of the explanation.

The differences in the stage of T cells, in vitro versus in vivo , overexpression versus deficiency, and the length of Tcf7 overexpression or deficiency could all contribute to the observed differences in the two studies. Although the discrepancy raises concern of our approach, results from our perturbation study on the effect of Bach2 on Prdm1 , Prdm1 on Tcf7 and Id2 on Id3 are all consistent with previous reports 17 , 36 , 37 , suggesting the validity of our in vitro assay in most cases.

S10 , which are considered as central memory T-cell precursors with high proliferative potential 38 , Another mechanism is by stimulating T-cell proliferation. We showed that overexpression of Bach2 enhances IL-2 driven T-cell proliferation in vitro and recall proliferation in vitro and in vivo. Our study further sheds light on the mechanisms by which Bach2 promotes T-cell proliferation. In the regulatory motif, Bach2 promotes Id3 expression but suppresses Prdm1 expression through direct binding to their promoter regions Fig.

Thus, by promoting Id3 expression, Bach2 stimulates T-cell proliferation. Prdm1 is known to antagonize Bcl6 , which promotes cell cycle by suppressing the expression of cell cycle inhibitor p27kip Consistently, we show that overexpression of Bach2 promotes Bcl6 expression but inhibits p27kip expression Fig.

Thus, Bach2 also stimulates T-cell proliferation by suppressing Prdm1 expression. All raw image files were reprocessed to normalize the data using R programme with a gcRMA method. Gene expression data was used to construct the regulatory network with the putative TFs using a reverse-engineering algorithm CLR Among the 1, putative TFs identified according to the domain predictions of protein sequences 18 , 1, were manually mapped to the latest mouse genome.

These 60 candidates were filtered by removing those whose knockout does not have any immune system phenotype as defined in MGI Mice were used at 8—16 weeks of age. Single-cell suspensions were prepared from spleens and mediastinal draining lymph nodes DLN , peripheral blood and lung. Red blood cells in the spleen, blood and lung were lysed with red blood cell lysis buffer Gibco and the cells were washed with complete RPMI. About 0. On the second day, the culture was replaced with fresh medium.

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On the third day, supernatant was collected and filtered through a 0. Cells were analysed for memory phenotype by flow cytometry on day 7. The numbers and phenotype of 2C T cells were analysed by flow cytometry 4 and 6 days later. To generate in vivo memory T cells, activated and transduced 2C T cells were adoptively transferred into B6 recipients.

Twentythree days later, the frequency, phenotype and function of persisting 2C T cells were analysed by flow cytometry. Twelve weeks later, cells were collected from spleen and analysed for 2C T-cell percentage and phenotype. A Millipore ChIP kit was used for chromatin immunoprecipitation. The membrane was stripped and re-blotted with the rabbit anti-mouse HRP-conjugated anti-Gapdh antibody Cell Signaling Technology for protein loading control. P -values for MRA and promoter enrichment results were calculated with a binomial test.

How to cite this article: Hu, G. Goldrath, A.

Selecting and maintaining a diverse T-cell repertoire. Nature , — Kaech, S. Molecular and functional profiling of memory CD8 T cell differentiation. Cell , — Wherry, E. Immunity 27 , — Intlekofer, A. Pearce, E. Science , — Sullivan, B. Antigen-driven effector CD8 T cell function regulated by T-bet. Natl Acad. USA , — Gattinoni, L. Zhao, D. Constitutive activation of Wnt signaling favors generation of memory CD8 T cells. Zhou, X. Immunity 33 , — Schober, S. Expression of the transcription factor lung Kruppel-like factor is regulated by cytokines and correlates with survival of memory T cells in vitro and in vivo.

Bai, A. Kruppel-like factor 2 controls T cell trafficking by activating L-selectin CD62L and sphingosinephosphate receptor 1 transcription. Carlson, C. Kruppel-like factor 2 regulates thymocyte and T-cell migration. Yang, C. Cannarile, M. Ji, Y. Kallies, A. Immunity 31 , — Rutishauser, R. Gray, P. Mouse brain organization revealed through direct genome-scale TF expression analysis. Faith, J. Large-scale mapping and validation of Escherichia coli transcriptional regulation from a compendium of expression profiles. PLoS Biol. Eppig, J. Group MGD. Nucleic Acids Res. Angelosanto, J.

Generating diversity: transcriptional regulation of effector and memory CD8 T-cell differentiation. D'Cruz, L. Joshi, N. Shen, C. Loss of IL-7R and ILR expression is associated with disappearance of memory T cells in respiratory tract following influenza infection. Hettmann, T. Teixeiro, E. Alon, U. Network motifs: theory and experimental approaches. Bernard, P. Acquisition of SOX transcription factor specificity through protein-protein interaction, modulation of Wnt signalling and post-translational modification.

Cell Biol. Sinner, D. Ikushima, H. Cell Stem Cell 5 , — Kuwahara, M. Sanjabi, S. Masson, F. Ochiai, K.