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. 2021 Jul 26;27(9):1530–1535. doi: 10.1038/s41591-021-01464-w

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© The Author(s) 2021

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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Extended Data Fig. 5 — (a) To characterize spike- and SEB-reactive CD4 T cells regarding their single or combined expression of the cytokines IFN-γ, IL-2, and TNF-α, CD4 T cells positive for combined expression of CD69 and IFN-γ were divided into four subpopulations according to additional expression of IL-2 and TNF-α. Using NOT Boolean Gating, all CD4 T cells which were not CD69 + IFN-γ + were analyzed for CD69 + IL-2+ and CD69 + TNF-α + CD4 T cells. Using OR Boolean Gating, CD69 + IL-2+ and/or CD69 + TNF-α + CD4 T cells were divided into IL-2 single, TNF-α single or IL-2+TNF-α + cells. After subtraction of background reactivity from negative control stimulations, the sum of these 7 subpopulations was set to 100%. A similar strategy was applied for CD8 T cells. (b) Cytokine-expression profiles of spike-specific CD4 and CD8 T cells in all individuals showing single or combined expression of the cytokines IFN-γ, interleukin (IL) 2 and tumor necrosis factor (TNF) α. To allow for robust statistics, analysis was restricted to samples with at least 30 cytokine-positive T cells (n = 178 for CD4 and n = 112 for CD8 T cells). Numbers refer to the percentage of cells expressing the respective cytokine. IFN, interferon; IL, interleukin; TNF, tumor necrosis factor.