Figure 4
Figure 4. TLR-stimulated CDPs preferentially home to LNs in expense of the BM in steady-state hosts. CDPs were FACS-isolated from the BM of CD45.1+ and CD45.2+ mice. CD45.1+ CDPs were stimulated for 2 hours in cultures containing Flt3L and CpG, whereas CD45.2+ CDPs were cultured in Flt3L alone. After culture, CDPs were washed, mixed in a one-to-one ratio, CFSE-labeled, and adoptively transferred intravenously into CD45.1xCD45.2 F1 steady-state hosts. After 30 hours, BM, spleen, and subcutaneous LNs were analyzed. The transferred mix of CD45.1+ CpG-stimulated CDPs and CD45.2+ CDPs from control cultures is shown in panel A. Cells gated on NK1.1− CD19− are shown for the different organs after 30 hours in vivo (B left column). To better distinguish CDP-derived cells from the host, cells were gated CFSE+, and the ratio of CD45.1+ CpG-stimulated and CD45.2+-nonstimulated cells was determined (B middle column). The CD11c and MHC II expression of CD45RA− cells is shown for each subset (B right column). The results of 1 representative experiment of 3 independent experiments of each recipient mouse are shown.

TLR-stimulated CDPs preferentially home to LNs in expense of the BM in steady-state hosts. CDPs were FACS-isolated from the BM of CD45.1+ and CD45.2+ mice. CD45.1+ CDPs were stimulated for 2 hours in cultures containing Flt3L and CpG, whereas CD45.2+ CDPs were cultured in Flt3L alone. After culture, CDPs were washed, mixed in a one-to-one ratio, CFSE-labeled, and adoptively transferred intravenously into CD45.1xCD45.2 F1 steady-state hosts. After 30 hours, BM, spleen, and subcutaneous LNs were analyzed. The transferred mix of CD45.1+ CpG-stimulated CDPs and CD45.2+ CDPs from control cultures is shown in panel A. Cells gated on NK1.1 CD19 are shown for the different organs after 30 hours in vivo (B left column). To better distinguish CDP-derived cells from the host, cells were gated CFSE+, and the ratio of CD45.1+ CpG-stimulated and CD45.2+-nonstimulated cells was determined (B middle column). The CD11c and MHC II expression of CD45RA cells is shown for each subset (B right column). The results of 1 representative experiment of 3 independent experiments of each recipient mouse are shown.

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