Figure 4
From: Regulatory T cells are essential to promote proper CD4 T-cell priming upon mucosal infection
Dendritic cell (DC) trafficking from the vaginal mucosa is impaired in the absence of regulatory T cells (Tregs). Foxp3DTR or control mice were injected with diphtheria toxin (DT) and infected intravaginally with herpes simplex virus-2 (HSV-2) 186ΔKpn as above. (a) Expression of C-C motif chemokine ligand 21 (CCL21) in the draining lymph nodes (dLNs) of Treg-sufficient or Treg-depleted mice 2 days after infection determined by quantitative PCR analysis (relative to the housekeeping gene hypoxanthine-guanine phosphoribosyltransferase (HPRT)). (b) Fluorescein isothiocyanate (FITC) painting experimental layout. (c) Representative flow plots showing the fraction of CD11b+/CD8− DCs in the dLNs that stained positive for FITC 2 days after infection. Cells were first gated on major histocompatibility complex (MHC) II/CD11c+/CD11b+ and CD8− cells. (d) Representative data showing the number of FITC+ DCs that had entered the dLNs from the infected vaginal tract 2 days after the mice were infected. (e) Ex vivo antigen presentation experimental layout. (f) The amount of interferon-γ (IFNγ) produced by anti-HSV-2 CD4 T cells after 60 h of co-culture with DCs sorted from Treg-sufficient or Treg-depleted mice 2 days after infection. On the left, cells were cultured without addition of exogenous antigen, whereas, on the right, cells were provided HSV-2 ex vivo during culture. All data come from or are representative of at least two similar, independent experiments with 3–5 mice in each group that showed similar results. Each data set was first screened for outliers using the ROUT test with a Q value of 0.2%. Statistical significance was then determined using an ordinary one-way analysis of variance followed by Tukey’s test to compare the mean of each group with the mean of all other groups.*P⩽0.05, **P⩽0.01, ***P⩽0.001 ****P⩽0.0001. Error bars show s.d. WT, wild type.
