Project description:To determine the activation patterns of ILC2s and associated ILC2-intrinsic functional molecules triggered by the high fiber diet, we administered either a control or high fiber diet in WT mice and performed RNA sequencing of FACS-sorted ILC2s from mouse colons. RNAseq libraries were prepared from 1,000 sorted colonic lamina propria ILC2s (CD45+Lin-CD90.2+CD127+KLRG1+) by the Epigenomics Core at WCM using the Clontech SMARTer® Ultra® Low Input RNA Kit V4 (Clontech Laboratories). Libraries were sequenced on an Illumina HiSeq 2500, generating 50 bp single-end reads. Two samples from two control diet-fed WT SPF mice and two samples from two high fiber diet-fed WT SPF mice were used.
Project description:To investigate the comprehensive mRNA expression profile of ILC2s from IPF patients, we performed bulk RNA-sequencing analysis of ILC2s sorted from IPF patients and healthy controls.
Project description:To investigate the transcriptional regulation of Xbp1s in ILC2s, sorted large intestinal ILC2s from mice were used for CUT&Tag sequencing to analyze the target of Xbp1s.
Project description:Group 2 innate lymphoid cells (ILC2s) play critical roles in allergic lung inflammation through initiating and amplifying type 2 immune responses. However, the molecular mechanisms underlying pathogenic ILC2 activation remain largely unknown. Here, we show that lung ILC2s exhibit increased mTORC1 activation in allergic asthma. Genetic ablation of RAPTOR, an adaptor protein of the mTORC1 complex, results in reduced IL-5 and IL-13 production in ILC2s and protects mice from allergic inflammation. Pharmacological inhibition of mTORC1 by rapamycin also suppresses ILC2 pathogenic activation and ameliorates allergic lung inflammation. Mechanistically, mTORC1 signaling promotes ILC2 activation through metabolic regulation of epigenetics to maintain neurointerin U receptor 1 (NMUR1) expression, which mediates neural-ILC2 interaction via the NMU-NMUR1 axis. These findings identify mTORC1 as a novel regulator to control the neural-ILC2 interaction and highlight mTORC1 as a potential therapeutic target for allergic asthma.
Project description:RNA sequencing was performed for the DCA- or vehicle-treated ILC2s sorted from the large intestine of C57BL/6 mice (n = 2 per group).
Project description:RNA sequencing was performed for the toyocamycin- or vehicle-treated ILC2s sorted from the large intestine of C57BL/6 mice (n = 3 per group).
Project description:The colonic lamina propria contains a distinct population of Foxp3+ T regulatory cells (Tregs) that modulate responses to commensal microbes. Analysis of gene expression revealed that the transcriptome of colonic Tregs is distinct from splenic and other tissue Tregs. Rorγ and Helios in colonic Tregs mark distinct populations: Rorγ+Helios- or Rorγ-Helios+ Tregs. We uncovered an unanticipated role for Rorγ, a transcription factor generally considered to be antagonistic to Foxp3. Rorγ in colonic Tregs accounts for a small but specific part of the colon-specific Treg signature. (1) Total colonic and splenic Foxp3+ Treg comparison: Lymphocytes were isolated from colonic lamina propria and spleens of Foxp3-ires-GFP mice, where GFP reports Foxp3 expression. TCRb+CD4+GFP+ cells were double sorted into Trizol. (2) Colonic Rorγ+ and Rorγ- Treg comparison: Foxp3-ires-Thy1.1 reporter mice were crossed to Rorc-GFP reporter mice to generate mice that report both Foxp3 and Rorγ expression. Rorγ+Foxp3+ Tregs (TCRb+CD4+Thy1.1+GFP+) and Rorγ-Foxp3+ Tregs (TCRb+CD4+Thy1.1+GFP-) from colonic lamina propria were double sorted into Trizol.To reduce variability and increase cell number, cells from multiple mice were pooled for sorting and at least three replicates were generated for all groups. RNA from 1.5-3.0 x104 cells was amplified, labeled and hybridized to Affymetrix Mouse Gene 1.0 ST Arrays.
