TLR7 controls myeloid-derived suppressor cells expansion and function in the lung of C57BL6 mice infected with Schistosoma japonicum
Fig 4
Effect of TLR7 on the phenotype and function of MDSCs in the lung during S. japonicum infection.
(A-D) C57BL/6, TLR7-/- mice were infected with 40 ± snail eggs through the skin, and lung single-cell suspensions were isolated from naive and infected WT and TLR7-KO mice after 6–7 weeks for flow cytometry. (A) The proportions of the MDSCs subtypes in the lungs were evaluated by flow cytometry. Representative results (left) and the statistical graph (right) are shown. Data are expressed as the mean ± SD of 3–6 mice. (B) Phenotypic analysis of CD11b+Gr1+ cells from the lungs of mice by different fluorescence-labeled Abs to mouse surface markers, including PD-L1, PD-L2, CD115, CD135, and CD103. Representative results (up) and statistical graph (down) are shown. Data are expressed as the mean ± SD of 3–9 mice. (C) Single cell suspensions of lung cells from mice were stimulated with PMA and ionomycin. The expression of GM-CSF, IFN-γ, IL-1α, IL-6 and IL-10 were detected in MDSCs by FACS analysis. Data are expressed as the mean ± SD of 3–7 mice. A representative of two independent experiments is shown. (D) MDSCs were harvested from the lung and the ROS level was evaluated by flow cytometry. CD11b+Gr1+ cells were gated and the percentage of CM-H2DCFDA+ cells is shown as the mean ± SD of 6 samples pooled from three independent experiments. Data are expressed as the mean ± SD of 3 mice. (A-D) * p < 0.05, ** p < 0.01 compared with the corresponding control, unpaired t-test was used.