Figure 5. Tissue resident CD301b+ DCs activate TH2 cells during bladder infection.
a, Bladders were collected at 0h or 12h after infection for staining of CD301b (green), uroepithelium (red) and nuclei (blue). White arrow indicates CD301b+ DC infiltration into epithelium. White bar marks 50 μm. The number of CD301b+ cells per field was counted, the ratio between the number of CD301b+ cells in epithelium and the number of those cells in lamina propria (LP) was determined. n=18 field, and each group has 6 mice. b, Samples were collected on Day 0 and Day 1 after infection for flow cytometry analysis. The population shown was pre-gated on 7AAD-CD45+ MHC class II+ single cells and n=7 mice per group. c, Bladders from WT 4get mice with DT, CD301bDTR 4get mice with PBS and CD301bDTR 4get mice with DT were collected on Day 3 after infection for flow cytometry analysis. The quantification of IL-4+ percentage was shown, n=8 mice. d, Flow cytometry analysis after second infection was shown. n=6 mice. e, Bladders from WT Great mice with DT, CD301bDTR Great mice with PBS and CD301bDTR Great mice with DT were collected on Day 3 after infection for flow cytometry analysis. The quantification of IFN-γ+ percentage was shown, n=6 mice. f, Flow cytometry analysis after second infection was shown, n=5 mice. g, Three days after the first or second infection, bladders from WT and CD301bDTR mice were collected for superficial BEC staining. White bar represents 25 μm. MFI of superficial BECs was quantified (right), n=6 mice. h, Three days after the first or second infection, bladders from WT and CD301bDTR mice were collected to determine CFU count. Data are shown as mean ± SD and were analyzed by an unpaired two-tail t test (a,b), an ordinary one-way (c-f) or two-way (g,h) ANOVA with a Turkey’s multiple comparison post-test. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001, ns=not significant.