Figure 5
Activation of A2A and A2B receptors downregulates T-cell activation during acute toxoplasmosis. C57BL/6 mice were inoculated orally with 40 cysts of ME-49. (a) At day 9 after infection, TCRβ+ CD4+ lymphocytes from spleen, MLN, LP, or IEL were cell-sorted. mRNA was extracted and quantitative RT-PCR of A2A and A2B receptors mRNA was performed. Data are pooled from four independent experiments. Graph bars represent the median with interquartile range. (b) At day 3 p.i., mice were treated daily with the combination of A2A and A2B receptor agonists or DMSO until day 9. Spleen, MLN, LP, and IEL cells were isolated and restimulated with PMA and ionomycin in the presence of Brefeldin A. Cells were stained for TCRβ, CD4, α4β7, CCR9, Ki-67, IFN-γ, and Foxp3 and analyzed by flow cytometry. Data are pooled from three independent experiments (n=14). The non-parametric Mann–Whitney U-test was used to determine statistical differences (*P<0.05, **P<0.01, ***P<0.001). DMSO, dimethyl sulfoxide; IEL, intraepithelial lymphocyte; LP, lamina propria; MLN, mesenteric lymph node; RT-PCR, reverse transcription-PCR.
