Figure 1
Intestinal CD4+ Foxp3− T lymphocytes have a high capacity to generate adenosine, in a CD73-dependent manner. (a, b) Cells were isolated from spleen, MLNs, LP, and epithelium of the small intestine (IEL) of C57BL/6, stained for TCRβ, CD4, CD39, CD73, Foxp3 expression and analyzed by flow cytometry. (a) Data are expressed as dot plots of CD39 or CD73 and Foxp3 expression on TCRβ+ CD4+-gated cells or (b) as the level of expression (mean fluorescence intensity, MFI) of CD39 or CD73 by CD73+ cells, among CD4+ Foxp3− or CD4+ Foxp3+ T lymphocytes. Data are representative of four independent experiments (n=10). The non-parametric Kruskal–Wallis test followed by selected comparison by Dunn’s multiple comparison tests was used. (c) CD4+GFP− and CD4+GFP+ T lymphocytes were isolated from the spleen, MLN, and LP of WT or CD73KO-Foxp3eGFP mice. Lymphocytes were incubated with AMP and the supernatants were tested for adenosine content by HPLC. Data are pooled from four independent experiments. Graph bars represent the median with interquartile range. (d) Amounts of adenosine were assessed by HPLC in sera from WT or CD73 mice. Data are pooled from three independent experiments (n=13). The non-parametric Mann–Whitney U-test was used. (*P<0.05, **P<0.01, ***P<0.001). GFP, green fluorescent protein; HPLC, high-performance liquid chromatography; LP, lamina propria; MLN, mesenteric lymph node; WT, wild type.
