Figure 5: S. aureus sustains CD11b⁺ M2 AM generation from non-resident monocyte.

(a,b) Quantitative RT–PCR analysis of the expression of Arg1, Fizz1, Ym1, IL-10, TGF-β (a) and iNOS (b) mRNAs in AMs from WT mice at the indicated time points after S. aureus priming. (c) AMs from WT mice or Tlr2^−/− mice at day 3 after S. aureus priming were co-cultured with naïve splenocytes and stimulated by Con A; 3 days later, [³H]-thymidine incorporation in T cells was analysed. (d) Quantitative RT–PCR analysis of the expression of Arg1, Fizz1, Ym1, IL-10 and TGF-β mRNAs in CD11b⁻ and CD11b⁺ AMs from WT mice at day 3 after S. aureus priming. (e) CD11b⁻ and CD11b⁺ AMs from WT mice or Tlr2^−/− mice at day 3 after S. aureus priming were co-cultured with naïve splenocytes and stimulated by Con A; 3 days later, the expression of Ki67 in T cells was evaluated. (f) Two days after CL₂MDP-lip treatment, WT mice were primed with S. aureus, and then CD11b⁻ or CD11b⁺ AMs from S. aureus-primed WT mice were adoptively transferred i.n. into these mice 3 days later. Survival of recipient mice was measured after infecting with 0.5 HA of PR8 immediately after adoptive transfer. (g) Six hours after sublethal irradiation, 5 × 10⁶ EGFP⁺ BM cells were adoptively transferred i.v. into WT mice and then primed with S. aureus 7 days later. The percentage of EGFP⁺ AMs in recipient mice at day 3 after S. aureus priming was evaluated by flow cytometry. Two-tailed Student’s t-tests; NS, not significant; *P<0.05, **P<0.01, ***P<0.001. Data are expressed as mean±s.e.m. Data represent three independent experiments with at least five mice per group in (a,b), or represent two independent experiments with at least five mice per group in (f,g), or represent three independent experiments with three wells per treatment in (c,e).