Notch signaling licenses allergic airway inflammation by promoting Th2 cell lymph node egress
Irma Tindemans, … , Ralph Stadhouders, Rudi W. Hendriks
Irma Tindemans, … , Ralph Stadhouders, Rudi W. Hendriks
Published April 7, 2020
Citation Information: J Clin Invest. 2020;130(7):3576-3591. https://doi.org/10.1172/JCI128310.
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Research Article Immunology Pulmonology

Notch signaling licenses allergic airway inflammation by promoting Th2 cell lymph node egress

Abstract

Allergic asthma is mediated by Th2 responses to inhaled allergens. Although previous experiments indicated that Notch signaling activates expression of the key Th2 transcription factor Gata3, it remains controversial how Notch promotes allergic airway inflammation. Here we show that T cell–specific Notch deficiency in mice prevented house dust mite–driven eosinophilic airway inflammation and significantly reduced Th2 cytokine production, serum IgE levels, and airway hyperreactivity. However, transgenic Gata3 overexpression in Notch-deficient T cells only partially rescued this phenotype. We found that Notch signaling was not required for T cell proliferation or Th2 polarization. Instead, Notch-deficient in vitro–polarized Th2 cells showed reduced accumulation in the lungs upon in vivo transfer and allergen challenge, as Notch-deficient Th2 cells were retained in the lung-draining lymph nodes. Transcriptome analyses and sequential adoptive transfer experiments revealed that while Notch-deficient lymph node Th2 cells established competence for lung migration, they failed to upregulate sphingosine-1-phosphate receptor 1 (S1PR1) and its critical upstream transcriptional activator Krüppel-like factor 2 (KLF2). As this KLF2/S1PR1 axis represents the essential cell-intrinsic regulator of T cell lymph node egress, we conclude that the druggable Notch signaling pathway licenses the Th2 response in allergic airway inflammation via promoting lymph node egress.

Authors

Irma Tindemans, Anne van Schoonhoven, Alex KleinJan, Marjolein J.W. de Bruijn, Melanie Lukkes, Menno van Nimwegen, Anouk van den Branden, Ingrid M. Bergen, Odilia B.J. Corneth, Wilfred F.J. van IJcken, Ralph Stadhouders, Rudi W. Hendriks

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Figure 7

Notch signals promote Th2 cell lymph node egress via transcriptional activation of the KLF2/S1PR1 axis.

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Notch signals promote Th2 cell lymph node egress via transcriptional act...
(A) Gene set enrichment analyses using the preranked differentially expressed (DE) genes shown in Figure 6C. (B) Expression levels of selected genes (n = 3–4 biological replicates; error bars denote SEM). **P < 0.01; ***P < 0.001; adjusted P values from DESeq2. (C) Flow cytometry analysis of surface CCR4, CCR8, and CXCR3 expression on WT and Notch-deficient Th2 cells from MedLNs and lungs of mice treated with HDM and OVA. (D) PCA using expression values of the 681 DE genes (Figure 6C) from MedLN WT and Notch-deficient OTII Th2 cells as well as Gata3+ MedLN and BAL Th2 cells from GATIR mice treated with HDM (as in Figure 1A). The 1-dimensional side plot illustrates how PC2 clusters WT with OTII MedLN cells and WT BAL Th2 cells with N1N2ΔCD4/ΔCD4 OTII MedLN Th2 cells. (E) Expression fold changes for indicated genes in Gata3+ Th2 cells from MedLNs versus BAL or WT versus Notch-deficient OTII transgenic MedLN Th2 cells (representative of 3 experiments; color code as in D). (F) RBPJκ ChIP-Seq signal (from the 8946 T-ALL cell line; ref. 51) in the Dtx1 and Gata3 (both canonical Notch target genes) and the S1pr1 and Klf2 loci. (G) Quantitative PCR measurements of Notch ligand genes in populations of CD11b+ migratory DCs, lymphoid endothelial cells (LECs), and fibroblastic reticular cells (FRCs) from the MedLNs of mice 3 days after PBS or HDM exposure.