Fig. 6

Alveolar macrophages are required for the protective effects of ADNVs. (A-G) Simplified scheme of AMs depletion experiment. C57BL/6 mice (n = 5) were intratracheally instilled with 100 µL CL₂MDP (5 mg/mL) for 2 days. The mice were then treated with LPS (1 mg/kg, i.t.), followed by ADNVs administration (25 mg/kg, i.v.). The animals were sacrificed 24 h and subjected to functional analysis (A); (B) Flow cytometry of CD11c⁺SiglecF⁺ AMs gated at CD45⁺CD11b⁻ CD64⁺; (C) Flow cytometry analysis of MMs (CD45⁺CD11b⁺ F4/80⁺); (D) Total cell counts, and (E) Protein concentration in BALF; (F) H&E staining of lung tissues and lung injury score. Scale bar, 200 μm; (G) qPCR of the indicated cytokines in lung tissues. (H-M) The simplified scheme of macrophage adoptive transfer experiment. Murine bone marrow-derived macrophages (BMDMs) were prepared and pre-conditioned with or without ADNVs for 2 h. The mice (n = 5) were subjected to LPS (1 mg/kg) challenge, 4 h later, the cells were then adoptively transferred to mice that were pre-depleted pulmonary macrophages. (I) H&E staining of lung tissues and lung injury score (Scale bar, 200 μm); (J) Total cell counts, (K) Protein concentration, and (L) the ratios of macrophages and neutrophils in BALF; (M) qPCR of the indicated cytokines in lungs tissues. Shown are representative images and the data are expressed as the mean ± SD, *P < 0.05, **P < 0.01, ***P < 0.001