Induction of IL-10-producing type 2 innate lymphoid cells by allergen immunotherapy is associated with clinical response

•A thorough characterization of IL-10-producing type 2 innate lymphoid cell (ILC2s)•IL-10-producing ILC2s restore epithelial integrity and suppress Th2 responses•Symptom severity inversely correlates with after immunotherapy•IL-10-producing play a critical role in tolerance induction to aeroallergens The immune cells allergen immunotherapy that confers the sensitizing is unclear. Here, we report interleukin-10-producing (IL-10+ ILC2s) modulating grass-pollen allergy. We demonstrate KLRG1+ but not KLRG1– ILC2 produced IL-10 upon activation IL-33 retinoic acid. These attenuated Th responses maintained integrity. IL-10+ were lower patients allergy when compared healthy subjects. In prospective, double-blind, placebo-controlled trial, demonstrated competence produce was restored who received sublingual immunotherapy. underpinning mechanisms associated modification retinol metabolic pathway, cytokine-cytokine receptor interaction, JAK-STAT signaling pathways ILCs. 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GATA-3 transcription factor expression induced both 1C 1D), confirmed phenotype. transdifferentiating IFN-γ-producing (Bal exposed IL-1β, transforming growth β (TGF-β) IL-12, respectively. none conditions led 1B). Analyzing CRTH2 CD127+ ILCs, observed previously lacked 2A). four populations (KLRG1+CRTH2+; KLRG1–CRTH2+; KLRG1–CRTH2–) IL-33, found IL-10, whereas restricted any 2B). KLRG1+CRTH2+, KLRG1–CRTH2+ 2C). observation ILC2. able IL-10. To rule out possibility KLRG1+CRTH2+ consequence preferential outgrowth contaminating generated clones KLRG1–CRTH2+, using OP9-DL1 cloning efficiency 9%–16%. All derived IL-13. Fifty percent 25% substantially contrast, minor (6%) confirm KLRG1+CRTH2+/− 2D). Taken together, belong lineage precursors. then patterns genes ILC2s. single-cell Cellular Indexing Transcriptomes Epitopes Sequencing (scCITE-seq) revealed that, stimulation, upregulated CTLA4 IL2RA IL-10–KLRG1+ GATA3 PTGDR2 protein differentially expressed between IL-10– 3A). Expression other FOXP3 could detected. Using qRT-PCR, encoding molecules characterize (Treg) cells. (CD25) transcripts enhanced 3B 3C), HELIOS, BLIMP1, CCR4, CCR8, IRF4 same 3B). asked helped shape downstream tissue environment. address this, co-cultured days. proliferation rate measured. IL-5-producing IL-2 plus IL-7 expanded decreased IL-4+ latter neutralized anti-IL-10. blocked Th17 well and, similarly, this mediated IL-5+IL-10– IFN-γ indicating 3D–3F). partially prevented phosphorylation CD28 diminished addition IL-10-blocking antibody S2) affect inhibiting (Taylor 2007Taylor Joss Akkoç Wenig Daigle Flory Blaser ICOS costimulations via src homology domain-containing tyrosine phosphatase 1.J. 76-83Abstract (87) attenuate IL-10-dependent manner, stimulate present 2-mediated airways, i.e., polyp chronic rhinosinusitis (CRSwNP) S3). help maintain disrupted allergen. IL10R epithelium stimulation triggers 4A). Next, set cultures air-liquid interface system 4B). Fully differentiated ciliated goblet 48 h polarized production. followed exposure (Phleum pratense, Phlp) additional h. included sample first Phlp subsequently measurements transepithelial electrical resistance (TEER) amplified deterioration 4C). co-culture disintegration Addition IL-10-neutralizing protective restoring That restores supported junction zonula occludens-1 (ZO-1) downregulated (Vroling 2008Vroling Jonker M.J. Luiten Breit T.M. Primary house dust mite extract shows activated individuals.Am. Respir. Cell Mol. Biol. 38: 293-299Crossref (40) 2015Golebski Egmond E.J. Roschmann K.I. EGR-1 DUSP-1 important negative regulators pro-allergic epithelium.Mol. 65: 43-50Crossref (10) permanently state factors (TFs) nuclear κB (NF-κB) (i.e., p65) AP-1 c-Myc) family being baseline volunteers. At time, TFs known inhibit EGR-1) reduced prompted us investigate effects constitutively Figure 4D demonstrates manner pro-inflammatory NF-κB (NFKB1) (MYC) anti-inflammatory EGR1, 4D). Dysregulated may impact integrity, IL-6 IL-8 implicated promoting th