GCNT1-mediated O-glycosylation of the sialomucin CD43 is a sensitive indicator of notch signaling in activated T cells

Eric Perkey, Dave Maurice De Sousa, Leolene Carrington, Jooho Chung, Alexander Dils, David Granadier, Ute Koch, Freddy Radtke, Burkhard Ludewig, Bruce R. Blazar, Christian W. Siebel, Todd V. Brennan, Jeffrey Nolz, Nathalie Labrecque, Ivan Maillard

Research output: Contribution to journalArticlepeer-review

3 Scopus citations

Abstract

Notch signaling is emerging as a critical regulator of T cell activation and function. However, there is no reliable cell surface indicator of Notch signaling across activated T cell subsets. In this study, we show that Notch signals induce upregulated expression of the Gcnt1 glycosyltransferase gene in T cells mediating graft-versus-host disease after allogeneic bone marrow transplantation in mice. To determine if Gcnt1-mediated O-glycosylation could be used as a Notch signaling reporter, we quantified the core-2 O-glycoform of CD43 in multiple T cell subsets during graft-versus-host disease. Pharmacological blockade of Delta-like Notch ligands abrogated core-2 O-glycosylation in a dose-dependent manner after allogeneic bone marrow transplantation, both in donor-derived CD4+ and CD8+ effector T cells and in Foxp3+ regulatory T cells. CD43 core-2 O-glycosylation depended on cell-intrinsic canonical Notch signals and identified CD4+ and CD8+ T cells with high cytokine-producing ability. Gcnt1-deficient T cells still drove lethal alloreactivity, showing that core-2 O-glycosylation predicted, but did not cause, Notchdependent T cell pathogenicity. Using core-2 O-glycosylation as a marker of Notch signaling, we identified Ccl19-Cre+ fibroblastic stromal cells as critical sources of Delta-like ligands in graft-versus-host responses irrespective of conditioning intensity. Core-2 O-glycosylation also reported Notch signaling in CD8+ T cell responses to dendritic cell immunization, Listeria infection, and viral infection. Thus, we uncovered a role for Notch in controlling core-2 O-glycosylation and identified a cell surface marker to quantify Notch signals in multiple immunological contexts. Our findings will help refine our understanding of the regulation, cellular source, and timing of Notch signals in T cell immunity.

Original languageEnglish (US)
Pages (from-to)1674-1688
Number of pages15
JournalJournal of Immunology
Volume204
Issue number6
DOIs
StatePublished - Mar 15 2020

Bibliographical note

Funding Information:
This work was supported by the National Institute of Allergy and Infectious Diseases (R01-AI091627 to I.M. and R37-AI34495 to B.R.B.), the Canadian Institutes of

Funding Information:
B.R.B. receives remuneration as an advisor to Kamon Pharmaceuticals, Five Prime Therapeutics, Regeneron Pharmaceuticals, Magenta Therapeutics, and BlueRock Therapeutics as well as research support from Fate Therapeutics, RXi Pharmaceuticals, Alpine Immune Sciences, AbbVie, Blue-Rock Therapeutics, Leukemia & Lymphoma Society, Children’s Cancer Research Fund, and Kids First Fund. He is a cofounder of Tmunity. The other authors have no financial conflicts of interest.

Publisher Copyright:
© 2020 American Association of Immunologists. All rights reserved.

PubMed: MeSH publication types

  • Journal Article
  • Research Support, N.I.H., Extramural

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