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. 2020 Mar 7;69(7):1205–1216. doi: 10.1007/s00262-020-02535-6

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© The Author(s) 2020

Open AccessThis 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. 1 — Human CD19⁺ B cells and their BTK phosphorylation in response to ADO. a The expression of the ectonucleotidases CD39 and CD73 measured by FACS in PBMC of healthy donors (n = 60), PBMC of tumor patients (n = 42) and in TIL of HNSCC patients (n = 39). Data are shown as means ± SD. b RNA levels for the four adenosine receptors (ADOR) in isolated peripheral B cells of healthy donors (n = 10) and tumor patients (n = 5). c Human B cells were stimulated with anti-µ-F(ab’)₂ and stained for anti-p-BTK (pY223). The experiment was performed with gated B cells (n = 3) and isolated B cells in PBMC (n = 5). d Pre-incubation with ADO induced a decreased BTK phosphorylation in a concentration-dependent manner after stimulation with anti-µ-F(ab’)₂ (n = 3). e Incubation with ADO decreased the ability of B cells to phosphorylate BTK in a time-dependent manner. Results are expressed as percentage of phosphorylated BTK shown as mean ± SD (n = 10). f The ability of ADO to decrease BTK phosphorylation in peripheral B cells is comparable in B cells of healthy donors (n = 10) and HNSCC patients (n = 7). Results are expressed as percentage of phosphorylated BTK. The five-minute value was set to 100%. All data are shown as mean ± SD. not significant (ns); p < 0.05 (*); p < 0.01 (**); p < 0.001 (***); p < 0.0001 (****)