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. 2006 Nov 27;203(12):2649–2660. doi: 10.1084/jem.20060155

Figure 7.

Figure 7.

Characterization of mucosal T reg cells. (A–J) Representative staining profiles of surface CD4 expression and intracellular Foxp3 (B, E, and H) on pooled tracheal digest preparations from groups of ≥15 animals gated for CD25 from control animals (A and B) and sensitized animals exposed to a single (B and E) or multiple (G and H) OVA aerosols. Isotype controls are shown in (A, D, and G). Tracheal digest cells gated for Foxp3 from the same groups of animals express surface CD4 and CD25 as illustrated in C, F, and J, respectively. (K) Total number of Foxp3 cells per trachea in OVA-sensitized animals during repeated OVA aerosol exposure/rest/reexposure cycles. Data are representative (n ≥ 2) experiments at each time point and derived from intracellular staining profiles and total digest yields after accounting for isotype control staining. (L) Mean proliferative responses of LN cells obtained from OVA-sensitized animals in the presence or absence of CD4+CD25+ cells derived from the tracheal mucosa of sensitized animals exposed to multiple OVA aerosols. Data shown are from one representative experiment CD4+CD25+ were added at ratios of 1:10 (LN to CD4+CD25+, shaded bars) and 1:20 (checked bars). Addition of CD4+CD25+ cells from unexposed controls (1:10) was without effect (not shown). (M) Proliferative responses of LN CD4+ T cells from OVA/CFA-sensitized animals in the presence of OVA (50 μg/ml) and DCs sorted from tracheal digest of naive animals (▪) or animals sensitized and exposed to a single (◯) or multiple (•) OVA aerosols. Proliferation is shown as mean CPM of triplicate cultures and is representative of at least two individual experiments. (N) Proliferative responses of LN CD4+ T cells from OVA/CFA-sensitized animals in the presence of AMDCs sorted from animals sensitized and exposed to multiple OVA aerosol (no additional OVA added to culture). Proliferation in L, M, and N is shown as mean CPM of triplicate cultures and is representative of two or three experiments in each case. *, P < 0.001; **, P < 0.0005.