Fig. 3. Direct and indirect contact with allogeneic ASCs enhances the activity of CD39 and CD73 in Tregs leading to increased degradation of proinflammatory eATP and eAMP.

a The frequency of CD39⁺ cells within CD4⁺FoxP3⁺ T cell population and the intensity of CD39 expression (MFI, median fluorescence intensity) by CD4⁺FoxP3⁺CD39⁺ Tregs in the studied culture conditions (n = 9). b The frequency of CD73⁺ cells within CD4⁺FoxP3⁺ T cell population and the intensity of CD73 expression (MFI) by CD4⁺FoxP3⁺CD73⁺ Tregs in the indicated culture conditions (n = 9). *p < 0.05 for comparisons with Tregs from monocultures at day 7. **p < 0.05 for comparisons with Tregs from monocultures at day 14. c The intensity of degradation of eATP, eAMP and eADO by Tregs from the indicated culture conditions in relation to standard Treg monocultures (INDEX, n = 4). *p < 0.05 for comparisons between Treg monocultures and the given culture condition. **p < 0.05 for direct vs indirect cocultures. ***p < 0.05 for direct cocultures vs monocultures supplemented with ASC-derived SNs. ****p < 0.05 for indirect cocultures vs monocultures supplemented with ASC-derived SNs. d Correlations between eATP, eAMP and eADO degradation by Tregs from the indicated culture conditions and their inhibition of Tconv proliferation in 2:1, 1:1 and 1:2 Treg:Tconv ratios together (n = 48) were calculated with Spearman’s rank correlation, R and p values are given. Inhibition of Tconv proliferation was defined by the proliferation INDEX of Tconvs in the presence of Tregs. A lower proliferation INDEX of Tconvs in the presence of Tregs corresponds to a higher Treg suppressive potential. The differences between the culture conditions were calculated with a two-sided Mann–Whitney U test with correction. In all boxplots the median is indicated by the symbol within the box, lower and upper bounds of the boxes correspond with the 25th and 75th percentiles. The lower and upper whiskers indicate minimum and maximum values, respectively. MFI is presented as value to power of 3 or 4 (e.g. 1E4 = 1 × 10⁴). Source data are provided as a Source Data file.