Notch ligation by Delta1 inhibits peripheral immune responses to transplantation antigens by a CD8+ cell–dependent mechanism
Kenneth K. Wong, … , Jonathan R. Lamb, Margaret J. Dallman
Kenneth K. Wong, … , Jonathan R. Lamb, Margaret J. Dallman
Published December 1, 2003
Citation Information: J Clin Invest. 2003;112(11):1741-1750. https://doi.org/10.1172/JCI18020.
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Article Immunology

Notch ligation by Delta1 inhibits peripheral immune responses to transplantation antigens by a CD8+ cell–dependent mechanism

Abstract

Notch signaling plays a fundamental role in determining the outcome of differentiation processes in many tissues. Notch signaling has been implicated in T versus B cell lineage commitment, thymic differentiation, and bone marrow hematopoietic precursor renewal and differentiation. Notch receptors and their ligands are also expressed on the surface of mature lymphocytes and APCs, but the effects of Notch signaling in the peripheral immune system remain poorly defined. The aim of the studies reported here was to investigate the effects of signaling through the Notch receptor using a ligand of the Delta-like family. We show that Notch ligation in the mature immune system markedly decreases responses to transplantation antigens. Constitutive expression of Delta-like 1 on alloantigen-bearing cells renders them nonimmunogenic and able to induce specific unresponsiveness to a challenge with the same alloantigen, even in the form of a cardiac allograft. These effects could be reversed by depletion of CD8+ cells at the time of transplantation. Ligation of Notch on splenic CD8+ cells results in a dramatic decrease in IFN-γ with a concomitant enhancement of IL-10 production, suggesting that Notch signaling can alter the differentiation potential of CD8+ cells. These data implicate Notch signaling in regulation of peripheral immunity and suggest a novel approach for manipulating deleterious immune responses.

Authors

Kenneth K. Wong, Matthew J. Carpenter, Lesley L. Young, Susan J. Walker, Grahame McKenzie, Alyson J. Rust, George Ward, Laura Packwood, Karen Wahl, Luc Delriviere, Gerard Hoyne, Paul Gibbs, Brian R. Champion, Jonathan R. Lamb, Margaret J. Dallman

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Figure 2

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Dl1 is able to activate Notch signaling in reporter cell assays and in T...
Dl1 is able to activate Notch signaling in reporter cell assays and in T cells. (a) CHO/N2-luc cells, which contain a construct that reports Notch pathway activation by expression of luciferase, were cultured with mitomycin C–treated Kb, Kb/Dl1, Ab, or Ab/Dl1. Untransfected CHO cells were used to control cell numbers in the assay and do not activate the Notch pathway in CHO/N2-luc cells. Cell lysates were assayed for luciferase activity after 24 hours. (b) Quantitative RT-PCR was used to assess the expression of Notch receptors and Notch ligands by L cells. All samples were normalized by assessment amplification of 18s RNA. Mouse embryonic RNA (day 11) was used as a positive control in the experiment, and transcript levels in other samples are expressed relative to this. (c) Positively selected CD4+ or CD8+ cells were cultured in the presence of immobilized CD3 Ab (0.12 μg/ml), CD28 Ab (2 μg/ml), and immobilized Dl1-Fc fusion protein (40 μg/ml). After 4 hours, cells were harvested and RNA extracted. Samples were analyzed for Hes1 transcripts and normalized using 18s amplification by quantitative RT-PCR. Data are expressed as fold induction compared with cells incubated with CD3 and CD28 Ab’s in the absence of Dl1-Fc fusion protein. All data are representative of at least three repeat experiments.