Notch signaling in T cells is essential for allergic airway inflammation, but expression of the Notch ligands Jagged 1 and Jagged 2 on dendritic cells is dispensable

doi:10.1016/j.jaci.2016.11.046

Journal of Allergy and Clinical Immunology

Volume 140, Issue 4, October 2017, Pages 1079-1089

https://doi.org/10.1016/j.jaci.2016.11.046 Get rights and content

Background

Allergic asthma is characterized by a T_H2 response induced by dendritic cells (DCs) that present inhaled allergen. Although the mechanisms by which they instruct T_H2 differentiation are still poorly understood, expression of the Notch ligand Jagged on DCs has been implicated in this process.

Objective

We sought to establish whether Notch signaling induced by DCs is critical for house dust mite (HDM)–driven allergic airway inflammation (AAI) in vivo.

Methods

The induction of Notch ligand expression on DC subsets by HDM was quantified by using quantitative real-time PCR. We used an HDM-driven asthma mouse model to compare the capacity of Jagged 1 and Jagged 2 single- and double-deficient DCs to induce AAI. In addition, we studied AAI in mice with a T cell–specific deletion of recombination signal–binding protein for immunoglobulin Jκ region (RBPJκ), a downstream effector of Notch signaling.

Results

HDM exposure promoted expression of Jagged 1, but not Jagged 2, on DCs. In agreement with published findings, in vitro–differentiated and HDM-pulsed Jagged 1 and Jagged 2 double-deficient DCs lacked the capacity to induce AAI. However, after in vivo intranasal sensitization and challenge with HDM, DC-specific Jagged 1 or Jagged 2 single- or double-deficient mice had eosinophilic airway inflammation and a T_H2 cell activation phenotype that was not different from that in control littermates. In contrast, RBPJκ-deficient mice did not experience AAI and airway hyperreactivity.

Conclusion

Our results show that the Notch signaling pathway in T cells is crucial for the induction of T_H2-mediated AAI in an HDM-driven asthma model but that expression of Jagged 1 or Jagged 2 on DCs is not required.

Graphical abstract

Section snippets

Methods

For detailed methods, including mice used, experimental protocols and statistical analysis, see the Methods section in this article's Online Repository at www.jacionline.org.

Jagged 1 is upregulated on in vitro GM-CSF BMDCs on exposure to HDM

Because several research groups have shown a role for Jagged in the orchestration of T-cell responses by using GM-CSF BMDCs,6, 11, 14, 15, 16 we first investigated the expression of Notch ligands on BMDCs on stimulation with the pro-T_H2 stimulus HDM and the pro-T_H1 stimulus LPS. GM-CSF BMDCs were cultured from wild-type (WT) mice and sorted at day 9 into CD11c⁺MHC class II^intF4/80⁻CD115⁺ GM–monocyte-derived dendritic cells (MoDCs), CD11c⁺MHCII^highF4/80⁻CD115⁻ GM-DCs, and CD11c⁺MHCII^intF4/80⁺

Discussion

Notch signaling in T cells is crucial to induce a T_H2 response. This was shown earlier in mouse models using parasite antigens4, 7 and in asthma models using OVA.⁸ In line with these reports, we found that mice with T cell–specific RBPJκ deficiency did not mount a T_H2 response in an HDM-induced mouse AAI model. However, the role of the Notch ligands Jagged 1 and Jagged 2 in T_H2 induction remains more elusive. Here we show that on HDM exposure, Jagged 1 is specifically upregulated on migratory

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Taken together, these effects on the airway epithelium could be seen as a bonus of SAHM1 topical therapy in the airways targeting inflammatory cells. Nevertheless, our previous finding that HDM-driven airway inflammation is diminished in mice with conditional RBPj deletion exclusively in the T-cell lineage demonstrated the critical role of Notch signaling in T cells.12 Our study has implications for the therapeutic use of peptidomimetic compounds in general.
The Notch signaling pathway has been implicated in the pathogenesis of allergic airway inflammation. Targeting the active Notch transactivation complex by using the cell-permeable, hydrocarbon-stapled synthetic peptide stapled α-helical peptide derived from mastermind-like 1 (SAHM1) resulted in genome-wide suppression of Notch-activated genes in leukemic cells and other models. However, the efficacy of SAHM1 in allergic asthma models has remained unexplored.
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Topical therapeutic intervention with SAHM1 or a control peptide was performed during sensitization, challenge, or both with HDM in mice. Airway inflammation was assessed by using multicolor flow cytometry, and bronchial hyperreactivity was studied. Additionally, SAHM1 therapy was investigated in mice with established allergic airway inflammation and in a model in which we neutralized IFN-γ during HDM challenge to support the T_H2 response and exacerbate asthma.
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: Supported in part by Lung Foundation Netherlands grants 3.2.12.087 and 3.2.12.067.

: Disclosure of potential conflict of interest: I. Tindemans receives grant support from the Lung Foundation of The Netherlands; travel support from Lung Foundation of The Netherlands. NRS of The Netherlands, and NVVI of The Netherlands. M. Lukkes, B. W. Li, and R. W. Hendriks receives grant support from the Lung Foundation of The Netherlands. The rest of the authors declare that they have no relevant conflicts of interest.

^∗: These authors contributed equally to this work.

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Mechanisms of allergy/immunologyNotch signaling in T cells is essential for allergic airway inflammation, but expression of the Notch ligands Jagged 1 and Jagged 2 on dendritic cells is dispensable

Background

Objective

Methods

Results

Conclusion

Graphical abstract

Section snippets

Methods

Jagged 1 is upregulated on in vitro GM-CSF BMDCs on exposure to HDM

Discussion

J Biol Chem

Immunity

Immunity

Cell

Trends Immunol

Immunobiology

Immunity

Immunity

Immunity

Immunity

Immunity

Immunity

J Allergy Clin Immunol

Immunity

Immunity

Immunity

Inflammatory dendritic cells—not basophils—are necessary and sufficient for induction of Th2 immunity to inhaled house dust mite allergen

J Exp Med

Dendritic cell and epithelial cell interactions at the origin of murine asthma

Ann Am Thorac Soc

Mechanisms of allergy/immunology
Notch signaling in T cells is essential for allergic airway inflammation, but expression of the Notch ligands Jagged 1 and Jagged 2 on dendritic cells is dispensable