Figure 1.

Increased secretion of Th2-associated cytokines in SOCS2^−/− mice in vitro and in vivo. (A) CD4⁺CD25⁻ splenocytes purified from WT and SOCS2^−/− mice were cultured for 3 or 5 d with 2 µg/ml of plate-bound anti-CD3, 1 µg/ml of plate-bound anti-CD28, and 5 U/ml IL-2 in triplicate and cytokine production was evaluated by ELISA. Data are representative of at least three independent experiments. (B) Naive CD4⁺CD25⁻CD44⁻ splenocytes were cultured for 7 d under Th2 polarization conditions. Frequency of CD4⁺IL-4⁺ cells (Th2 cells) was evaluated at days 4 and 7 by intracellular staining. Data are representative of three independent experiments performed in triplicate. (C) SOCS2^−/− and WT mice (n = 5 per group) were i.p. injected with 10,000 S. mansoni eggs or PBS. 10 d later, splenocytes were restimulated with anti-CD3 and anti-CD28 or SEA and cytokine production was measured by ELISA. (D and E) The footpads of WT and SOCS2^−/− mice (n = 5 per group; D) or RAG-1^−/− mice reconstituted with CD4⁺ T cells sorted from WT (n = 4 per reconstituted group; E) or SOCS2^−/− mice (n = 7 per reconstituted group; E) were injected with 5,000 S. mansoni eggs or PBS. 10 d later, the amount of Th2 cells in popliteal LNs was determinate by IL-4 intracellular staining (D) or T1/ST2 (IL-33 receptor) staining (E). Data are representative of at least three experiments and presented as the mean and SEM of each group. *, P < 0.05; **, P < 0.01; ***, P < 0.001.