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IL18 signaling controls the CCR6-CCL20-dependent migration axis used by IL18R+ Tregs to enter the thymus

ABSTRACT: We identified transcriptional changes in Tregs in response to IL18 stimulation, including upregulation of CCR6

ORGANISM(S): Mus musculus

PROVIDER: GSE153153 | GEO | 2020/07/27

REPOSITORIES: GEO

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IL18 signaling controls the CCR6-CCL20-dependent migration axis used by IL18R+ Tregs to enter the thymus

Project description:IL18 signaling controls the CCR6-CCL20-dependent migration axis used by IL18R+ Tregs to enter the thymus

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IL18 signaling promotes homing of mature Tregs into the thymus

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A novel model to study deficiency of RORγt+ Tregs reveals net suppressive function and dependency on the CCR6/CCL20 axis for expansion and activation

Project description:Background: Recently, Tregs expressing the Th17 characteristic transcription factor RORγt were identified (biTregs). Both, pro- and anti-inflammatory functions have been described and the sum effect of biTreg deficiency in inflammation remains unknown. Furthermore, it is unclear, whether biTregs are specialized for the control of a defined type of immune response, as in particular Th2 or Th17 immunity. Methods: FACsorted CD4+ T-cells from fluorescence reporter mice, containing or lacking biTregs, were transferred into RAG1-/- recipients and the nephrotoxic nephritis model of glomerulonephritis was studied. Interactions of biTregs with various leukocyte populations were assessed by multicolor immunofluorescence. Furthermore, biTregs were compared to other Treg subtypes including suppression assays and RNAseq analyses. Finally, the role of the CCR6/CCL20 axis for biTreg biology was characterized. Results: Lack of endogenous biTregs resulted in aggravation of glomerulonephritis, demonstrating net immunosuppressive functions. However, contrary to our hypothesis, biTreg deficiency did not result in selectively overshooting Th2 or Th17 responses. Comprehensive analysis instead revealed a unique functional and transcriptional biTreg profile, including high production of IL-10 and dependency on IL-6R, cMAF and IKZF3 activation. Interestingly, we found that expression of the chemokine receptor CCR6 marked a particularly activated biTreg subset, whose absence significantly worsened glomerulonephritis. Unexpectedly, we found that signaling via the CCR6/CCL20 axis mediates biTreg expansion, rather than trafficking. Our data indicate net immunosuppressive effects of endogenous biTregs, with no preference for regulation of Th2 or Th17 responses. Surprisingly, signaling via the Conclusion: CCR6/CCL20 axis was integral for biTreg expansion and activation, which offers novel therapeutic options.

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Bulk RNA-seq of Tregs from WT mice and Ccr6 KO mice

Project description:The experiment was designed to compare transcriptomic differences between WT and Ccr6 KO Tregs during activation. WT and Ccr6 KO Tregs, cells were isolated from mice and cultured in vitro for 3 days with activation using anti-CD3/CD28 beads. Total RNA was extracted using the Trizol method. Quantity and quality were assessed using a Thermo Scientific™ NanoDrop™ 2000/2000c Spectrophotometer. Novogene Corporation Inc prepared the RNA-seq 250-300 bp insert cDNA library. Illumina HiSeq platform PE150 sequencing was used for sequencing, yielding 20M raw reads/sample. Mus Musculus mm39 was used as the reference genome for alignment.

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Gene expression analysis of the hippocampus in Il18+/+ and Il18-/- with restraint stress

Project description:Interleukin-18 (IL-18) was one of the inflammatory cytokines and related to major depressive disorder (MDD) and dementia. We previously investigated IL-18 knockout (Il18−/−) mice developed impaired learning and memory, more anxiety, and less motivation comparing to Il18+/+ mice. In the Il18−/− mice with acute stress, both the immobility time in the forced swim test and the tail suspension test were decreased, even though no difference was observed in the Il18+/+ mice. To reveal the mechanism, RNA-Seq was performed.

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Gene expression analysis of the hippocampus in Il18+/+ and Il18-/- mice at 12 weeks of age

Project description:Interleukin-18 (IL-18) was one of the inflammatory cytokines and related to major depressive disorder (MDD) and dementia. Compared with Il18+/+ mice, IL-18 knockout (Il18−/−) mice developed impaired learning and memory, more anxiety, and less motivation. To reveal the mechanism, the microarray was performed.

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IL18 signaling promotes homing of mature Tregs into the thymus

Project description:This SuperSeries is composed of the SubSeries listed below.

2020-07-27 | GSE153155 | GEO

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Project description:To detect the miRNA expression profile in CCR6+ regulatory T cells In this study, the total RNA was extracted from CCR6+ regulatory T cells and CCR6- regulatory T cells. Then, the expression profile of miRNA on these cells was detected by microarray.

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Gene expression analysis of the hippocampus in Il18+/+ and Il18-/- with restraint stress

Project description:Gene expression analysis of the hippocampus in Il18+/+ and Il18-/- with restraint stress

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