Figure 3.

miR-142-3p has different effects on CD4⁺CD25⁺ regulatory T cells and CD4⁺CD25⁻ T cells. (A) miR-142-3p inhibits the suppressor function of T_REG cells. CD4⁺CD25⁻ T cells (responder cells) were cultured alone, or co-cultured with T_REG cells, control oligo- or miR-142-3p- transfected T_REG cells in the presence of anti-CD3 and APCs. The proliferation (left) was determined by ³H-thymidine incorporation, and cytokines in the supernatants (right) were determined by ELISA. (B) Blockade of miR-142-3p impairs the activation of CD4⁺CD25⁻ T cells. Control CD4⁺CD25⁻ T cells, inhibitor control oligo- or miR-142-3p inhibitor-transfected CD4⁺CD25⁻ T cells were stimulated with anti-CD3 and irradiated APCs. The proliferation of cells (left) and the cytokines in the supernatants (right) were determined as above. (C) The blockade of miR-142-3p does not confer a suppressor phenotype on CD4⁺CD25⁻ T cells. CFSE-labelled native CD4⁺CD25⁻ T cells (responder) were cultured alone (group ‘none'), or co-cultured with T_REG cells, miR-142-3p inhibitor- or inhibitor control oligo-transfected CD4⁺CD25⁻ T cells at a ratio of 1:1, and stimulated with anti-CD3 and irradiated APCs. The proliferation of responder cells was analysed by flow cytometry. The results are shown as the averages of proliferation index (left) and the representative dot plot of flow cytometric analysis (right). The data in this figure are the representatives of two independent experiments (n=6 per group in each experiment); ^*P<0.05 compared with T_REG or controls. APC, antigen-presenting cell; ELISA, enzyme linked immunosorbent assay; IL-2, interleukin 2; IFN-γ, interferon-γ; T_REG, regulatory T cells.