Fig. 6
IL-12 supplementation rescues the defect in parasite control in CD11c-Cre OTUB1fl/fl mice. a Experimental design scheme of IL-12 administration and analysis. Mice were i.p. injected with either PBS or 150 ng IL-12 daily from 0 to 3 days p.i. b The parasite load in peritoneal cells was determined by semiquantitative PCR of tissue DNA after infection with 50,000 tachyzoites for 4 days (n = 8 for each group). c Percentages of infected peritoneal CD45+ cells were analyzed by flow cytometry at day 4 after infection with 50,000 tachyzoites. Representative flow cytometry plots (left panel) and statistics (right panel) are shown (n = 8 for each group). d, e Percentages of infected peritoneal leukocyte subpopulations (d) and DC subsets (e) were analyzed by flow cytometry at day 4 after infection with 50,000 tachyzoites (n = 8 for each group). f Mice were i.p. infected with five cysts of the DX strain. At day 30 p.i., the parasite loads in the brains of surviving mice were determined by semiquantitative PCR. Two independent experiments with comparable results were performed. One representative experiment is shown (n = 4 for each group). g Intracerebral parasites in the brains of T. gondii-infected OTUB1fl/fl and CD11c-Cre OTUB1fl/fl mice at 30 days p.i. A PBS-treated OTUB1fl/fl mouse (top left) shows a single focus consisting of a few T. gondii cysts and some tachyzoites in the white matter of the frontal lobe. The brain of a PBS-treated CD11c-Cre OTUB1fl/fl mouse (top right) shows an increased number of parasitic foci (arrows) with T. gondii cysts and tachyzoites in the white matter of the frontal lobe. In both an OTUB1fl/fl and a CD11c-Cre OTUB1fl/fl mouse, IL-12 application reduced the intracerebral parasitic load, with only single cysts scattered throughout the frontal lobe (arrows). Immunohistochemistry with polyclonal rabbit anti-T. gondii (BioGenex, Fremont, CA, USA) and slight counterstaining with hemalum; original magnification ×200; scale bar (a–d): 50 µm. The photomicrographs shown are representative of three mice per experimental group. Similar results were obtained in a second independent experiment. h The survival of the mice was monitored daily up to 30 days after infection with cysts of strain 5 DX (n = 8 for each group). Data are displayed as the mean ± SD. *p < 0.05, and ***p < 0.001; n.s. not significant
