1. The Temporal and Spatial Dynamics of Foxp3+ Treg Cell–Mediated Suppression during Contact Hypersensitivity Responses in a Murine Model 
Sari Lehtimäki, Terhi Savinko, Katharina Lahl, Tim Sparwasser, Henrik Wolff, Antti Lauerma, Harri Alenius, Nanna Fyhrquist 
Journal of Investigative Dermatology 
Volume 132, Issue 12, Pages (December 2012)
DOI: /jid
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2. Figure 1
Effects of forkhead box P3 (Foxp3+) cell depletion during sensitization phase. (a) Mice were treated with diphtheria toxin (DT) 1 day before sensitization. (b) The percentages of Foxp3+ cells among CD4+ T cells were determined in the draining lymph nodes (dLNs) by flow cytometry at various time points after sensitization and challenge. (c) Representative images of hematoxylin and eosin (H&E)–stained ear tissue of vehicle-treated and oxazolone-sensitized and challenged wild-type (WT) and depletion of regulatory T cell (DEREG) mice. Scale bar=100μm. (d) Ear swelling, (e) cellular infiltration, and (f) expression of several cytokines were significantly upregulated in DEREG mice compared with WT. *P<0.05; **P<0.01; ***P<0.001; bars represent mean±SD. HPF, high-power field; OXA, oxazolone (4-ethoxymethylene-2-phenyl-2-oxazolin-5-one); RU, relative units; sensDT, DT treatment prior to sensitization group; VEH, vehicle.
Journal of Investigative Dermatology  , DOI: ( /jid )
Copyright © 2012 The Society for Investigative Dermatology, Inc Terms and Conditions

3. Figure 2
Effects of forkhead box P3 (Foxp3+) cell depletion during elicitation phase. (a) Mice were treated with diphtheria toxin (DT) one day before elicitation. (b) The percentages of Foxp3+ cells were determined in the draining lymph nodes (dLNs) by flow cytometry at various time points after sensitization and challenge. (c) Representative images of hematoxylin and eosin (H&E)–stained ear tissue of vehicle-treated and oxazolone-sensitized and challenged wild-type (WT) and depletion of regulatory T cell (DEREG) mice. Scale bar=100μm. (d) At 24 hours after challenge, the ear swelling response, (e) cellular infiltration, and (f) expression levels of several cytokines were all at levels similar to those of the WT mice. HPF, high-power field; elicDT, DT treatment prior to elicitation group; OXA, oxazolone (4-ethoxymethylene-2-phenyl-2-oxazolin-5-one); RU, relative units; VEH, vehicle.
Journal of Investigative Dermatology  , DOI: ( /jid )
Copyright © 2012 The Society for Investigative Dermatology, Inc Terms and Conditions

4. Figure 3
Resolution of inflammation was impaired after forkhead box P3+ (Foxp3+) cell depletion. (a) Ear swelling response peaked at 24 hours after challenge in wild type (WT) mice (black dots), but was sustained at a high level both in the group depleted of Foxp3+ cells before sensitization (sensDT depletion of regulatory T cell (DEREG), white squares) and in the group depleted of Foxp3+ cells before elicitation (elicDT DEREG, gray triangles) at 48 hours and 96 hours after challenge. (b) The mRNA expression levels of several cytokines were sustained at a significantly higher level in DEREG mice compared with WT. (c) T-cell infiltration in the skin increased throughout the observation period in all groups. (d) The percentage of Foxp3+ cells increased in the skin of WT mice as a function of time, whereas the proportion remained low in both DEREG groups. (e) In sensDT DEREG mice, the percentage of Foxp3+ cells reached WT levels by 96 hours after challenge. *P<0.05; **P<0.01; ***P<0.001; bars represent mean±SD. HPF, high-power field; IHC, immunohistochemistry; RU, relative units.
Journal of Investigative Dermatology  , DOI: ( /jid )
Copyright © 2012 The Society for Investigative Dermatology, Inc Terms and Conditions

5. Figure 4
Increased percentages of T cells proliferated and expressed activated phenotype after forkhead box P3+ (Foxp3+) cell depletion. (a) CD4+ and CD8+ T-cell proliferation at different time points after challenge was assessed by Ki-67 staining and flow cytometric analysis. (b) The activation status of T cells was determined by analyzing the intracellular expression of CTLA-4 with flow cytometry. *P<0.05; **P<0.01; ***P<0.001; bars represent mean±SD. DEREG, depletion of regulatory T cell; elicDT, DT treatment prior to elicitation group; sensDT, DT treatment prior to sensitization group; WT, wild type.
Journal of Investigative Dermatology  , DOI: ( /jid )
Copyright © 2012 The Society for Investigative Dermatology, Inc Terms and Conditions

6. Figure 5
The expression of co-stimulatory molecules was enhanced in dendritic cells (DCs). (a) Flow cytometric analysis of CD11c+ DCs in the draining lymph nodes (dLNs) revealed upregulated expression of costimulatory molecules CD80 and CD86 in depletion of regulatory T cell (DEREG) mice compared with wild type (WT). (b) The mRNA expression of CD80 and CD86 in the skin was elevated in sensDT DEREG mice compared with WT at all-time points studied after challenge. Instead, in elicDT DEREG mice, the expression was higher compared with WT only at 48 hours and 96 hours after challenge. *P<0.05; **P<0.01; ***P<0.001; bars represent mean±SD; RU, relative units.
Journal of Investigative Dermatology  , DOI: ( /jid )
Copyright © 2012 The Society for Investigative Dermatology, Inc Terms and Conditions