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. Author manuscript; available in PMC: 2009 Jan 22.
Published in final edited form as: J Immunol. 2006 May 1;176(9):5329–5337. doi: 10.4049/jimmunol.176.9.5329

FIGURE 5.

FIGURE 5

PI-TReg cells from RAG−/− mice are suppressive in vitro and respond to Ag by IL-10 production. Spleen cells from (A) RAG+/+ and (B) RAG−/− mice were stained with FITC-anti-CD4 and PE-anti-CD25 and analyzed by FACS. The boxed area denotes CD4−CD25high cells constituting 1.1% of RAG+/+ and 0.05% of RAG−/− spleen cells. C–E, RAG−/− mice were rendered tolerant by 10 i.n. doses of Ac1-9[4Y]. Spleen cells were harvested 3 days after the final treatment and CD4+ T cells purified. CD4+ T cells were cultured at 5 × 104 cells per well with 1 × 105 cells per well of irradiated CD4-depleted splenocytes in vitro with 10 μg/ml Ac1-9 unless otherwise indicated. After 3 days, supernatants were harvested for cytokine ELISA or cultures pulsed with tritiated thymidine. The results are representative of groups of at least three mice per treatment group, and the experiment was repeated three times with similar results. Error bars indicate the SEM of triplicate cultures.