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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. 4 — a Volcano plot of the immune subsets percentage changes. b Left, Percentages of late transitional B cells (CD19⁺CD27¯CD24⁺⁺CD38⁺⁺) among living singlets. Middle, Normalized to baseline. Right, Representative cytometry plots of CD24 and CD38 expression among CD19⁺CD27¯ B cells. c Schematic of analyzing IL-10-producing B cells. d Percentages of IL-10⁺TNF¯ cells or IL-10⁺TNF¯/ IL-10¯TNF⁺ cell ratios among living B-cell singlets following stimulation. e Representative cytometry plots of B-cell intracellular cytokine expression. f, g Normalized MSI of ex-vivo expression of indicated markers. h Kinome tree showing a significant change (8 h vs. baseline) in VAP’s PBMC depleted of CD4⁺ T-cells. The nodes showing a Median Final Score (representing significance) >1.2 are marked with label(s). The Median Kinase Statistic represents the direction of effect. i, j Volcano plots of the immune subset percentage changes (left). Right panel of i, percentages of pDC (CD11c^¯CD123⁺) among living singlets. Upper right in j, Normalized percentages of CD4⁺ TEM (CD27^¯CD45RA^¯). Lower right in j, Representative cytometry plots of CD27 and CD45RA expression among CD4⁺ T cells. Expression of CD40LG (k) or IL10RA (l) transcripts in Th2 cells (n = 14 independent individuals). Data represent mean ± SEM (b, i). P-value was determined by paired two-tailed Mann-Whitney test in a, b, i, j or by paired two-tailed Student t test in d, f, g, k, l. q-values were generated using the two-stage step-up method (Benjamini, Krieger, and Yekutieli). For h, a mixed model statistical analysis was based on a one-tailed permutation test without multiple comparison correction (see Methods). ns, non-significant (p > 0.05). Each line in b, d, f, g, j and k, l links different time points of the same individual. In a, i, j, significantly enhanced or decreased immune subsets (p < =0.05) are marked. MSI, median signal intensity; ncMono, non-classical monocytes; pDC, plasmacytoid DC; CD8⁺ Treg, CD8⁺ regulatory T cells; TEM, effector memory T cells. For all the panels, HC, healthy controls, n = 10 independent individuals except for d and h (n = 9); PAP, pollen allergy patients, n = 16 independent individuals (no PAP in d and h); VAP, venom allergy patients, n = 18 independent individuals except for d and h (n = 9). Source data are provided as a Source Data file. Created in BioRender. Demczuk, A. (2024) BioRender.com/n34j539 (panel c).