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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. 2 — A–F Frequency of immune cells in control and HDM patch tests from reactive vs. non-reactive patients, measured by flow cytometry. The number in the graph indicates the percentage of cells in the positive gate. CR control patch, reactive patient, HR HDM patch, reactive patient, CNR control patch, non-reactive patient, HNR HDM patch, non-reactive patient. Representative examples. A, B CD3+ T lymphocytes, D, E CD207/CD1a positive LCs. C, F Fold changes (FC) in the percentage of detected immune cells between HDM patch test and control patch test from patients with irritant, non-reactive and reactive reactions to HDM. G Correlations between fold changes in the percentage of CD3+ T cells and LCs. Pearson correlation coefficient is shown. H Immunofluorescence staining of HDM-reactive patch test site. Inserts show the indicated optical fields at the epidermis (top) and in the dermis (bottom). Hub structures of co-localising CD207 (green) and CD3 (red) in the dermis. Epidermal layer stained with multi-cytokeratin (blue). DAPI stain for nuclei (grey). Scale bars: 500 μm, 50 μm (insets). A representative of n = 3 individual donors. I Functional assessment of skin barrier: TEWL measurements across patient groups. J Number of irritant (IR), non-reactive (NR) and reactive (R) cases with loss of function (LoF) variants in FLG compared to wildtype (WT). K Percentage of CD3+ T cells in control patch test sites identified by flow cytometry. Statistical significance was assessed by t-test. C, G NR n = 11, R n = 10, F, K NR n = 11, R n = 11, I, J IRR n = 4, NR n = 12, R n = 11. Statistical significance was assessed by the Kruskal–Wallis test with post hoc Dunn test (C, F, I) and unpaired ANOVA with post hoc Fisher test (K) following the normality Kolmogorov–Smirnov test of data distribution. Source data are provided as a Source Data file.