Notch promotes survival of pre–T cells at the β-selection checkpoint by regulating cellular metabolism

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Abstract

Notch signals are necessary for the functional outcomes of T cell receptor β-selection, including differentiation, proliferation and rescue from apoptosis. The mechanism underlying this requirement for T cell development is unknown. Here we show that Notch receptor and Delta-like 1 ligand interactions promoted the survival of CD4CD8 pre–T cells through the maintenance of cell size, glucose uptake and metabolism. Furthermore, the trophic effects of Notch signaling were mediated by the pathway of phosphatidylinositol-3-OH kinase and the kinase Akt, such that expression of active Atk overcame the requirement for Notch in β-selection. Collectively, our results demonstrate involvement of Notch receptor–ligand interactions in the regulation of cellular metabolism, thus enabling the autonomous signaling capacity of the pre–T cell receptor complex.

*Note: In the version of this article initially published online, in the fourth sentence of the abstract, the term "Atk" was a misspelling; this should be "Akt." In the fourth sentence of the second paragraph of the introduction, the name of the second kinase mentioned, "PI(3)K-dependent kinase 1," was incorrect; this should read "phosphoinositide-dependent kinase 1." These errors have been corrected for the HTML and print versions of the article.

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Figure 1: Requirement for Notch receptor and Delta-like 1 ligand interaction for TCR β-selection and pre–T cell survival.
Figure 2: Rag2−/− DN3 cells undergo apoptosis after withdrawal of Notch receptor–ligand interaction.
Figure 3: Rag2−/− DN3 cells undergo death and atrophy despite continued Bcl-2 expression.
Figure 4: Notch signals maintain glucose metabolism in pre–T cells.
Figure 5: Decreased Akt phosphorylation in the absence of Notch–Delta-like 1 ligand interactions.
Figure 6: Active Akt can restore the viability, cellular atrophy and decreased glycolytic rate of DN3 cells in the absence of Notch signaling.
Figure 7: Expression of active Akt restores the DN-to-DP transition in the absence of Notch–Delta-like 1 interaction.

Change history

  • 14 August 2005

    appended aop PDF with corrigendum (will be corrected for print issue), and placed footnote in XML at abstract

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Acknowledgements

We thank D.A. Vignali for the MIY retroviral vector; J. Maryanski for the MIG-TCRβ construct; W. Pear for the MigR retroviral vector; and G. Knowles for assistance in cell sorting. Supported by the Canadian Institutes of Health Research (Doctoral Research Award to M.C., and grant MOP 42387) and a Canada Research Chair in Developmental Immunology (J.C.Z.-P.).

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Correspondence to Juan Carlos Zúñiga-Pflücker.

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Supplementary information

Supplementary Fig. 1

The majority of Rag2−/− DN3 cells in OP9-DL1 and OP9-control cultures are non-cycling. (PDF 3241 kb)

Supplementary Fig. 2

Bcl-2 expression partially rescues the decline in cellularity following withdrawal of Notch-Delta-like-1 interaction. (PDF 1094 kb)

Supplementary Fig. 3

PI3K signals are indispensable for Notch-mediated trophic effects. (PDF 2995 kb)

Supplementary Table 1

Gene-specific primers used in RT-PCR analysis. (PDF 45 kb)

Supplementary Methods (PDF 42 kb)

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Ciofani, M., Zúñiga-Pflücker, J. Notch promotes survival of pre–T cells at the β-selection checkpoint by regulating cellular metabolism. Nat Immunol 6, 881–888 (2005). https://doi.org/10.1038/ni1234

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