Figure 5.

miR-155^−/− macrophages inhibit inflammatory cells and shape Th1/Th17 polarization (A,B) WT (n = 5) mice with dextran sulfate sodium (DSS) colitis were adoptive transferred with miR-155^−/− BMDMs or WT BMDMs (5 million/mouse) as schematic protocol indicated. The total number of LPMCs, CD4⁺ T cells, and CD8⁺ T cells of colon tissues at day 11 were counted. (C,D) miR-155^−/− mice (n = 5) with DSS colitis were i.p. injected with clodronate-liposomes (MDP) or control PBS-liposomes (PBS) as the schematic protocol indicated, and the total number of LPMCs, CD4⁺ T cells and CD8⁺ T cells of colon tissues at day 11 were counted. (E) OT-II mice splenocytes with incorporation of [³H]thymidine were cultured in the presence (+) or absence (−) of ConA, with (+) or without (−) miR-155 or WT BMDMs, then proliferation was assessed by [³H]thymidine incorporation. (F) CFSE-labeled naive OT-II T cells were cultured with OVA and miR-155^−/− or WT BMDMs culture supernatants for 4 days, and subjected to flow cytometry with gating of CD4⁺ T cells. (G) Flow cytometry of populations of CD4⁺IL-17⁺ T cells and CD4⁺IFN-γ⁺ T cells by OT-II T cells were primed as described in (F), then allowed to “rest” and re-stimulated for 6 h with anti-CD3 and anti-CD28. Numbers in outlined areas indicate percent cells in the gate. *P < 0.05, **P < 0.01 vs WT control [Student’s t-test in (B,D,E,G)]. Data are representative of three independent experiments [mean and SD in (A–D)]; n = 12–15 mice per group in (B,D). WT, wild-type.