Figure 2. IL-4Rα-expressing macrophages do not affect cytokine responses in granulomas.

Il4ra^−/lox, LysM^creIl4ra^−/lox, iLck^creIl4ra^−/lox and Il4ra^−/− mice were infected with 100 S. mansoni cercariae and killed 8 weeks later. Mesenteric lymph node (mLN) cells or liver granuloma-associated leukocytes (Liver) were then isolated and restimulated overnight with 20 µg/ml SEA before analyzed for ex vivo cytokine production capability. Staining of surface CD4 was followed by detection of IL-4, IL-10 and IFN-γ secretion in a cytokine catch assay or detection of intracellular levels of IL-13 after 4h of monensin treatment, as described in Methods. Live gate was placed on lymphocytes according to their forward- and side-scatter by flow cytometry. (A) Representative contour plots of IFN-γ, IL-4, IL-10 and IL-13-producing lymphocytes. Quadrant bars were set up on unstimulated cells and numbers represent percent cells in each quadrant. Data represent one out of three independent experiments with similar results. (B) Percentages of IFN-γ, IL-4, IL-10 or IL-13-producing cells were determined in gated CD4⁺ or non-CD4⁺ cell populations from mLN or liver granuloma-associated lymphocytes (Liver) (mean±SEM, n = 4). Data are representative of three independent experiments. *p<0.05; **p<0.001; ***p<0.001 compared to control Il4ra^−/lox mice.