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. 2014 Jan 2;10(1):e1003805. doi: 10.1371/journal.ppat.1003805

Figure 2. Production of IL-12 and IL-18 by Mtb-infected mϕ induce traditional markers of iNKT cell activation.

Figure 2

(A) iNKT cells were cultured either alone, with uninfected mϕ, H37Rv-infected mϕ, or αGalCer-loaded mϕ for 24 hours in the presence of blocking antibodies against IL-12p40 (20 µg/ml), IL-18 (10 µg/ml), CD1d (20 µg/ml), or respective isotype controls. Cells were stained for CD25 and mϕ were distinguished from iNKT cells by F4/80 staining. Supernatant was harvested at 24 hours and IFNγ measured by ELISA. (B) % reduction calculated as 100*[(iNKTH37Rv-mϕ−iNKTalone)−(iNKTAb+H37Rv-mϕ−iNKTalone)]/(iNKTH37Rv-mϕ−iNKTalone). Conditions with αGalCer stimulation calculated similarly. (C) iNKT cells were cultured with uninfected or H37Rv-infected WT or MyD88−/− mϕ for 24 hours. Cells were stained for CD25 and mϕ were distinguished from iNKT cells by F4/80 staining. Supernatant was harvested at 24 hours and IFNγ and IL-12p40 measured by ELISA. Error bars indicate mean ± SEM. **P<0.01 compared to isotype control. (One-way ANOVA with Dunnet's post-test). #P<0.05, ##P<0.01 compared to isotype control (data not shown) (unpaired Student's t-test). +P<0.05, ++P<0.01, +++P<0.001 (unpaired Student's t-test). Data are representative of, or compiled from, (A,B) three (anti-CD1d), four (anti-IL-12), two (anti-IL-18) and (C) two independent experiments.