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Series GSE203217 Query DataSets for GSE203217
Status Public on May 20, 2022
Title IL-33-activated calcitonin gene-related peptide-producing memory Th2 cells co-operate with somatosensory neurons to induce conjunctival itch
Organisms Homo sapiens; Mus musculus
Experiment type Expression profiling by high throughput sequencing
Summary Allergic conjunctivitis is a chronic inflammatory disease that is characterized by severe itch in the conjunctiva; but how neuro-immune interactions shape the pathogenesis of severe itch remains unclear. We identified a subset of memory-type pathogenic Th2 cells that preferentially expressed Il1rl1-encoding ST2 and Calca-encoding calcitonin gene-related peptide (CGRP) in the inflammatory conjunctiva using a single-cell analysis. The IL-33-ST2 axis in memory Th2 cells controlled the axonal elongation of the peripheral sensory C-fiber and the induction of severe itch. Pharmacological blockade and genetic deletion of CGRP signaling in vivo attenuated scratching behavior. The analysis of giant papillae from patients with severe allergic conjunctivitis revealed ectopic lymphoid structure formation with the accumulation of IL-33-producing epithelial cells and CGRP-producing pathogenic CD4+ T cells accompanied by peripheral nerve elongation. Thus, the IL-33-ST2-CGRP axis directs severe itch with neuro-reconstruction in the inflammatory conjunctiva and is a potential therapeutic target for severe itch in allergic conjunctivitis.
 
Overall design White blood cells of CD45+CD31- fractions that were freshly prepared from mice conjunctival tissue and sorted using a BD FACS Aria III (10,000 cells each, cell viability >98%) were encapsulated into droplets, and libraries were prepared using Chromium Single Cell 3’ Reagent Kits v3 according to manufacturer’s protocol (10X Genomics). The generated scRNA-seq libraries were sequenced using a 128 cycle (paired-end reads) with a NovaSeq 6000 (Illumina). Neuron Cells of CD45-CD31- and live cell+ fractions that were freshly prepared from trigeminal nerve tissue of mice and sorted using a BD FACS Aria III (10,000 cells each, cell viability >98%) were encapsulated into droplets, and libraries were prepared using Chromium Single Cell 3’ Reagent Kits v3 according to manufacturer’s protocol (10X Genomics). The generated scRNA-seq libraries were sequenced using a 128 cycle (paired-end reads) with a NovaSeq 6000 (Illumina). White blood cells of CD45+ and CD4+ fractions that were freshly prepared from Giant papillae from patients and sorted using a BD FACS Aria III (10,000 cells each, cell viability >98%) were encapsulated into droplets, and libraries were prepared using Chromium Single Cell 3’ Reagent Kits v3 according to manufacturer’s protocol (10X Genomics). The generated scRNA-seq libraries were sequenced using a 128 cycle (paired-end reads) with a NovaSeq 6000 (Illumina).
 
Contributor(s) Okano M, Hirahara K, Kiuchi M, Onoue M, Morimoto Y, Ikehara Y, Murakami A, Ebihara N, Nakayama T
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Submission date May 17, 2022
Last update date May 22, 2022
Contact name Masahiro Kiuchi
E-mail(s) aema2758@chiba-u.jp
Organization name Chiba univercity
Street address cyuoku inohana1-8-1
City chiba city
State/province chiba
ZIP/Postal code 2608670
Country Japan
 
Platforms (2)
GPL24247 Illumina NovaSeq 6000 (Mus musculus)
GPL24676 Illumina NovaSeq 6000 (Homo sapiens)
Samples (7)
GSM6164863 CD45+_conjunctival_tissue_Saline_mice
GSM6164864 CD45+_conjunctival_tissue_HDM_mice
GSM6164865 CD45+_PBMC_Human_patient
Relations
BioProject PRJNA839074

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Supplementary file Size Download File type/resource
GSE203217_RAW.tar 152.8 Mb (http)(custom) TAR (of MTX, TSV)
SRA Run SelectorHelp
Raw data are available in SRA
Processed data provided as supplementary file

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