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. 2023 May 19;14:2880. doi: 10.1038/s41467-023-38588-1

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© The Author(s) 2023

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 3 — Constellation-seq analysis enriched for 1161 transcripts in 2374 single T lymphocytes cells from patch test skin biopsies, n = 10 patients, 5 per group. A UMAP plot depicting clustering of T lymphocyte populations. B Cell subset defining markers (Wilcoxon rank test). C Number of T cell transcriptomes in control reactive (CR) and non-reactive (CNR) patients. The central line denotes the median, boxes represent the interquartile range (IQR), and whiskers show the distribution except for outliers. Outliers are all points outside 1.5 times the IQR. D Top transcription factors expressed in T cells from reactive (CR) and non-reactive (CNR) patients in the control samples. E Top biological pathways enriched at the control site in DEGs from patient reactive (CR) to HDM (KEGG database), p-value computed using the Fisher exact test, with Benjamini Hocheberg FDR correction. F UMAP plots showing expression of Th17 gene signature. G Th17 gene signature across patient groups, dot plot: size depicts % of expressing cells, colour intensity encodes mean expression in the group. H %IL17 producing CD3+ CD4+ T cells from PBMCs in irritant (I) non-reactive (NR) and reactive (R) patients. Kruskal–Wallis test with post hoc Dunn test. I A representative plot of IL17 expression in CD3+ CD4+ T cells I, NR and R patients. J Immunofluorescence staining of HDM-reactive patch test site. Inserts show the indicated optical field in the dermis. Hub structures of co-localising CD207 (blue), CD3 (green) and IL17 (red). Epidermal layer stained with multi-cytokeratin (blue). DAPI stain for nuclei (grey), Scale bars: 50 μm. Representative of n = 2. K) UMAP plots showing expression of TNF and TNFRSF1B across T cells (top) and APCs (bottom). L TNF and TNFRSF1B expression level across patient groups, dot plot: size depicts % of expressing cells, colour intensity encodes mean expression in the group. CR control reactive, HR HDM reactive, CNR control non-reactive, HNR HDM non-reactive. n = 5/group, C and H paired. Source data are provided as a Source Data file and via GEO.