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. 2021 Oct 8;12:5911. doi: 10.1038/s41467-021-26134-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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Fig. 3 — a Flow cytometric analysis of CD73 and CD39 expression on CD8 and CD4 T cells after activation of PBMCs (measurement on five time points from day 0 to day 7). Data were shown for four donors. b Schematic representation of CD73 released from CD8 T cells after PI-PLC treatment or activation. AMPase activity is measured by the conversion of 1,N⁶-etheno-AMP (eAMP) to 1,N⁶-etheno-ADO (eADO). c, d Determination of the AMPase activity of CD8 T cells and supernatants. 0.2 × 10⁶ CD8 T cells were treated with c PI-PLC (0.5 U/mL) or d stimulated with αCD3/αCD28 for 4 days. Histograms show CD73 expression of the cells at the time of eAMP incubation for one selected donor. The cells and supernatants were incubated with eAMP and degradation of eAMP to eADO was determined by HPLC (middle panel, mean of c six donors and d eight donors). The amount of soluble CD73 (sCD73) was measured by ELISA (right panel, mean of c six donors and d two donors). A two-tailed paired t-test was used to compare untreated and treated samples in c, d.