Figure 5.

In vivo, LPSs favoured mobilisation of a mo-DC like cell type in a TLR4-dependent fashion. 6–8 weeks-old C57BL/6J mice were injected intraperitoneally with 1xPBS (PBS), Bm-wt LPS (30 μg/mouse), Bm-wadC LPS (20 μg/mouse) or E. coli LPS (10 μg/mouse). Quantities of injected LPS, calculated by KDO weight ratio, ensured equal molar weight for each of them. 4, 12, 24 and 48 h post-injection, mice were sacrificed, single-splenocyte suspensions were prepared and analysed by flow cytometry. (A) Dot plots of CD11b and BST-2 staining and absolute numbers of the CD11b⁺BST-2^hi subset 12 h post-injection. (B) Absolute numbers of the CD11b⁺BST-2^hi subset in spleen from injected mice at indicated time-points. (C) Dot plots of DC-SIGN and CD64 staining of the pDC, cDC and CD11b⁺BST-2^hi subpopulations 12 h post-injection. (D) Dot plots of Ly6C and CD11b staining and absolute numbers of the CD11b⁺Ly6C^hi monocytes 12 h post-injection. (E) Dot plots of CD11c and MHCII staining of the CD11b⁺Ly6C^hi and CD11b⁺BST-2^hi subpopulations 12 h post-injection. Data displayed from (A to E) are representative of 3 independent experiments, n = 3 mice per condition in each experiment. (F) 6–8 weeks-old TLR4^+/+ or TLR4^−/− mice were injected with PBS, Bm-wt LPS, Bm-wadC LPS or E. coli LPS as described above. 12 h later, mice were sacrificed, single-splenocyte suspensions were prepared and total CD11c⁺ cells were analysed by flow cytometry. Dot plots of BST-2 and CD11b staining is representative of 3 independent experiments, n = 3 mice per condition in each experiment.