Fig. 3: BALB/c mice supplemented with IFN-γ display impaired resistance associated with strongly expanded Th2/1 hybrid responses.

a Experimental setup. BALB/c mice were infected with H. polygyrus at the age of 1.5 months and treated with recombinant IFN-γ (2.5 μg twice a day) from day 0 to 4 post infection. Mice were dissected at day 28 post infection. Graphs report the frequencies of Th2/1 hybrid and Th2 cells in CD4+ cells in blood over the course of infection. Data from two independent experiments are shown (n naïve = 3, infected ctr/ rIFN-γ treated = 6 mice) as mean ± SD. b Bar graphs depict the frequencies of Th2/1, Th2 and Th1 cells in siLP CD4+ T cells. Data from two independent experiments are pooled (n naïve = 4; infected ctr/ rIFN-γ treated = 6 mice). c Fecal egg counts determined over the course of infection. Data from two independent experiments are pooled (n = 6 mice/group) and shown as mean ± SD. d Adult worm counts at day 28 post infection. Data from two independent experiments are pooled (n = 6 mice/group). e Average number of eggs produced by individual female worms (typically eight worms per mouse, minimum 3) isolated from each mouse. Data from two independent experiments are combined (n infected ctr = 5; rIFN-γ treated = 3 mice). Note that expulsion was completed in one infected ctr and three rIFN-γ treated mice. f Granuloma count in small intestine at day 28 post infection. Data from two independent experiments are pooled (n = 6 mice/group). g Representative flow cytometry plots depicting T-bet and GATA-3 expression in CD4+ FoxP3− cells in spleen. Pie charts report the percentage of GATA-3+ T-bet+ Th2/1 cells in CD4+ GATA-3+ T cells isolated from the spleen at day 28. The size of the pie charts is adjusted according to the absolute cell count of GATA-3+ cells given above. Data from two independent experiments are pooled (n = 6 mice/group). h Representative flow cytometry plots depicting IFN-γ and GATA-3 expression of PMA/Ionomycin stimulated spleen cells at d28 post infection. Bar graph shows the frequencies of GATA-3+ IFN-γ+ cells in CD4+ FoxP3− cells isolated from spleen. Data from two independent experiments are pooled (n naïve = 4; infected ctr/rIFN-γ treated = 6 mice). Bars in b, d, e, f and h report the mean, circles represent individual mice. Statistical analysis was performed using unpaired t test or Mann–Whitney test in a, c, e and f and one-way ANOVA with Tukey’s multiple comparisons test in b and h. *p ≤ 0.05, **p ≤ 0.01, ****p ≤ 0.0001.