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Status |
Public on May 16, 2023 |
Title |
Transcriptomic analyses of cutaneous immune network in patiens with atopic dermatitis challenged with house dust mite (HDM) patch test |
Organism |
Homo sapiens |
Experiment type |
Expression profiling by high throughput sequencing
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Summary |
Our study investigates the activation and transcriptional programming of primary skin cells (FACS-purified Langerhans cells and single cell analysis of the whole biopsy) from patients with atopic dermatitis (n=22, nr = rensponding, nnr=nonresponding), exposed to a control or HDM patch test. 6 mm biopsies were taken from control and HDM patch test site 48h post application.
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Overall design |
Patients with moderate to severe atopic dermatitis eczema severity scores (EASI) median = 17.70, IQR:10.20 – 30.90, max = 51.40) under the care of a dermatologist in a tertiary referral centre were recruited to the study. 48h post application of a patch test to buttock skin, 11 out of the 28 patients were HDM-reactive, while In 12 patients, HDM did not induce a visible response, creating “HDM-non-reactive” group. 4 patients reacted to both HDM and control patch test "irritant", and one showed redness which was not patch test localised "undefined". 6 mm biopsies were taken from all patch test sites, and processed for flow cytometry or single cell RNA-seq analysis. CD207, CD1a, HLA-DR+ Langerhans cells (LCs)were FACS-sorted into RLT buffer and processed for bulk RNA-seq. Non-LCs cells were sorted and cryopreserved for further analysis with Constellation 10X. Freshly dissociated whole skin biopsies from 6 biopsies were suspended in RNAse-out buffer and processed on ice to the co-encapsulation of single cells with genetically-encoded beads (Drop-seq).
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Contributor(s) |
Polak ME, Sirvent-Bernal S, Vallejo AF |
Citation(s) |
37208336 |
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Submission date |
Sep 21, 2021 |
Last update date |
Aug 10, 2023 |
Contact name |
Marta Ewa Polak |
E-mail(s) |
m.e.polak@soton.ac.uk
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Phone |
2381205727
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Organization name |
University of Southampton
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Department |
Clinical and Experimental Sciences
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Lab |
Systems Immunology Group
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Street address |
Southampton General Hospital, Mailpoint 813, LE59,
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City |
Southampton |
State/province |
Hampshire |
ZIP/Postal code |
SO16 6YD |
Country |
United Kingdom |
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Platforms (3) |
GPL11154 |
Illumina HiSeq 2000 (Homo sapiens) |
GPL18573 |
Illumina NextSeq 500 (Homo sapiens) |
GPL20301 |
Illumina HiSeq 4000 (Homo sapiens) |
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Samples (61)
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Relations |
BioProject |
PRJNA764857 |
SRA |
SRP338039 |