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. 2024 Nov 26;15:10266. doi: 10.1038/s41467-024-54684-2

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© The Author(s) 2024

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 5 — a Relative frequency of FACS-sorted Th2 cells among living CD4⁺ T cells. q = 2.37e-4 (0 h), q = 1.24e-4 (8 h), q = 7.96e-4 (D1), q = 7.96e-4 (D7) – VAP vs HC; q = 0.0121 (0 h), q = 0.0303 (D1), q = 0.0121 (W2), q = 0.0303 (W6) and q = 0.0181 (W12) – PAP vs HC. b Pearson correlation between the sorted Th2 frequencies and CyTOF-measured Th2 percentages. c Heatmap of differentially-expressed genes (DEGs) between indicated time points in Th2. d Top-ranked GO processes among DEGs in c. e Left, Volcano plot highlighting DEGs in Th2. The substantially and significantly changed genes were labelled. Right, Schematic of IL-6 signaling. mRNA expression of SOCS3 (f), S1PR1 (g) and BCL3 (h) in Th2. P-values were adjusted with FDR. i Heatmap of 10 cytokines. j Serum IL-6 levels. Left, IL-6 levels among groups. q = 5.32e-3 (0 h), q = 9.6e-7 (8 h), q = 1.23e-5 (D1), q = 0.0206 (D7) – VAP vs HC. Normalized IL-6 levels (middle) or Scatter dot plots of absolute IL-6 levels (right). Mean for each time point in middle and right panels is labelled. k Confirmatory analysis of IL-6 levels by the NULISA platform. NPQ, NULISA protein quantification units (log2). Spearman correlation between serological IL-6 levels and cMono percentages among living singlets at baseline (l) or between the changed serological IL-6 levels and the altered percentages of cMono (m). Data represent mean ± SEM (a, j). P-value was determined by paired Bayes-moderated two-tailed t test [c, e, f-h] or non-paired [a, j (left)] two-tailed Mann-Whitney test, by paired two-tailed Student’s t test in b, j (middle), k, by two-tailed paired Wilcoxon signed-rank enrichment test (d), or by two-tailed exact permutation distribution test in l, m. q values were generated using the two-stage step-up method (Benjamini, Krieger, and Yekutieli). NS or unlabeled, not significant (p > 0.05). Each line in f-h and j, k links different samples of one individual. AIT, antigen-specific immunotherapy; APC, antigen-presenting cells; CyTOF, Mass cytometry; FACS, fluorescence-activated cell sorting. For all the panels, HC, healthy controls, n = 10 independent individuals except for panel k (n = 8); PAP, pollen allergy patients, n = 16 independent individuals; VAP, venom allergy patients, n = 18 independent individuals except for panels a-h, k (n = 15). Source data are provided as a Source Data file. Created in BioRender. Demczuk, A. (2024) BioRender.com/t00y783 (panel e).