Figure 3. Adoptive transfer of highly purified inflammatory monocytes can rescue antigen transport and CD4 T cell priming in DT treated CCR2-DTR mice.
CCR2-DTR mice were infected and treated with DT or PBS on days 7, 9 and 11 and received highly purified CD45.1+ IMs on days 8 and 10. (A) Mice were euthanized on day 12 post infection and flow cytometry was performed on lungs and mLNs to track the engraftment of adoptively transferred IMs. (B) The expression of cell surface markers CD11b, MHC II, CDIIc and CD103 by adoptively transferred IMs in the lung and mLN was determined. (C) CFU plots from day 12 mLNs of CCR2-DTR mice rescued with IMs. Dots marked in red represent mice that received double sorted IMs of greater than 99% purity. (D) CCR2-DTR mice received a dose of naive ESAT-6-specific C7 T cells the day before infection and were treated with DT on days 7, 9 and 11 and received purified IMs or PBS on days 8 and 10. Lungs were harvested on day 15 and ESAT-6-specific C7 T cells were visualized. (E) Cumulative data of experiment shown in (D) showing the total number of ESAT-6-specific C7 T cells in day 15 lungs. (F) CCR2-DTR mice were depleted on days 7, 9 and 11 and received CCR2 WT or CCR2 KO IMs on days 8 and 10. Day 12 mLNs were harvested for CFU counts. Each dot represents an individual mouse. Error bars denote SEM. Data are representative of two independent experiments.
Figure 3—figure supplement 1. Sorting of IMs from CCR2-GFP mice. FLT3-, cKit-, GFP+ cells were sorted from the bone marrow of CCR2-GFP mice.
