P47phox−/− Mice Are Compromised in Expansion and Activation of CD8+ T Cells and Susceptible to Trypanosoma cruzi Infection
Figure 3
In vitro functional activation of p47phox−/− macrophages in response to T. cruzi infection.
(A&B) Isolated splenocytes from WT and p47phox−/− mice were in vitro infected with live or heat-inactivated (HIA) T. cruzi for 24 h (± IFNγ). Cells incubated with phorbol myristate acetate (PMA) were used as positive controls. (A.a) H2DCFDA oxidation by intracellular ROS was monitored by fluorimetry. (A.b) Nitroblue tetrazolium (NBT) reduction to formazan blue crystals by cellular superoxide was visualized by light microscopy. (B) RT-PCR quantitation of iNOS mRNA level. (C) BM (panels a&c) and splenic (panels c&d) cells were stimulated with IFN-γ, washed, and then exposed to T. cruzi (live or heat-inactivated) for 24 h. The release of IFN-γ (panels a&b) and TNF-α (panels c&d) in supernatants was monitored by an ELISA. ND: not detectable.