Figure 2. Ebi3^−/−Il10^−/− T_regs that developed in a mixed bone marrow chimera can function in vitro and in vivo.

Congenically labeled wild type bone marrow and knock out bone marrow was mixed at a 1:1 ratio and injected into sub lethally irradiated Rag1^−/− mice. (A) After 8 weeks Thy.1.2⁺ wild type T_reg cells or knock out T_reg cells were purified by FACS from the bone marrow chimeric mice and cultured in the inserts of a Transwell™ plate in the presence of wild type T_conv cells and anti-CD3 and anti-CD28 coated latex beads. Wild type naive T_conv cells were activated in the presence of anti-CD3 and anti-CD28 coated beads in the bottom chamber of a Transwell™ for 72 h. Proliferation was determined by [³H]-thymidine incorporation. Data represents the mean± SEM of two independent experiments. (p-value; NS: Not significant). (B) Purified wild type or Ebi3^−/−Il10^−/− bone marrow chimera-derived T_regs were injected into Rag1^−/− mice in the presence of congenically marked naïve T_conv cells. The expansion of naïve Thy1.1 CD4⁺ T cells were assessed by flow cytometry. Data represents the mean ± SEM of two independent experiments with 3–4 mice per group (p-value *=0.06).