P47phox−/− Mice Are Compromised in Expansion and Activation of CD8+ T Cells and Susceptible to Trypanosoma cruzi Infection
Figure 5
In vivo ROS, nitric oxide, and cytokine profile in p47phox−/− mice infected with T. cruzi.
Mice (WT and p47phox−/−) were infected with T. cruzi as described in Materials and Methods. (A) Splenic single cell population from infected mice (30 dpi) were cyto-spinned on glass slides. Shown is fluorescence staining for dihydroethidium (measures intracellular ROS, binds DNA). Cells stained with DAPI (binds nuclei, blue) are shown as controls. (B) Splenocytes were harvested from T. cruzi-infected mice at day 30 pi, and in vitro incubated for 24 h (± TcL). Shown is intracellular nitric oxide (panel a) and nitrite levels in the supernatant (panel b), determined by using DAF-FM staining/fluorimetry and Griess reagent assay, respectively. (C) Splenocytes were harvested from infected mice at day 7, 14, 21, and 30 post-infection, and incubated in vitro (± TcL) for 48 h. Shown are the TNF-α (a&b), IFN-γ (c&d) and IL-10 (e&f) levels in cell-free supernatants, determined by an ELISA. ND: not detectable.