Skin Contact Irritation Conditions the Development and Severity of Allergic Contact Dermatitis Marlene Bonneville, Cyril Chavagnac, Marc Vocanson, Aurore.

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Skin Contact Irritation Conditions the Development and Severity of Allergic Contact Dermatitis Marlene Bonneville, Cyril Chavagnac, Marc Vocanson, Aurore Rozieres, Josette Benetiere, Ingrid Pernet, Alain Denis, Jean-Francois Nicolas, Ana Hennino Journal of Investigative Dermatology Volume 127, Issue 6, Pages 1430-1435 (June 2007) DOI: 10.1038/sj.jid.5700726 Copyright © 2007 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 1 Skin contact irritation is more severe in C57BL/6 mice than in BALB/C mice. A single skin painting with 0.1% (⋄), 0.3% (□) or 0.5% (△) DNFB on the left ear (and vehicle on the right ear) of (a) BALB/C (white) and (b) C57BL/6 (black) mice induced within hours an immediate ear skin inflammation. Results are expressed as the mean ear swelling±SD. (c–f) Histological analysis of ear sections from BALB/C and C57BL/6 mice, 6hours after vehicle (c, d) or DNFB (e, f) sensitization. Hematoxylin and eosin staining. Original magnification × 100. Results are representative of three independent experiments. Journal of Investigative Dermatology 2007 127, 1430-1435DOI: (10.1038/sj.jid.5700726) Copyright © 2007 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 2 The severity of ICD correlates with high levels of IL-1β/IL-6 mRNA and low levels of IL-10 mRNA. (a) Detection and quantification of mRNA for IL-1α, IL-1β, IL-6, and IL-10 by RNAse protection assay in the skin of DNFB-treated BALB/C mice 3hours after sensitization. Controls include expression of mRNA in the ear skin of naive mice and of vehicle-treated mice at 3hours. (b) Histograms representations of the relative quantities of IL-1β (black bars), IL-6 (white bars), and IL-10 (gray bars) mRNA recovered from the different experimental conditions, as compared to expression of glyceraldehyde-3-phosphate dehydrogenase mRNA as reference gene. (c, d) Comparison of the expression levels of (c) IL-1β and (d) IL-10 mRNA levels by RNase protection assay in the skin of BALB/C (white bars) and C57BL/6 (black bars) mice, 3 and 6hours following 0.3% DNFB sensitization. Controls include mRNA levels in the ear skin of naive mice and of vehicle-sensitized mice. mRNA relative quantities were compared with glyceraldehyde-3-phosphate dehydrogenase mRNA as standards. Journal of Investigative Dermatology 2007 127, 1430-1435DOI: (10.1038/sj.jid.5700726) Copyright © 2007 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 3 The intensity of ICD reaction dictates the severity of ACD responses. DNFB-induced skin inflammation was analyzed in groups of five (a) BALB/C and (b) C57BL/6 mice within hours (left panel) and days (right panel) following sensitization with DNFB 0.1% (⋄,♦), 0.3% (□,▪), or 0.5% (△,▴) on the left ear and vehicle on the controlateral ear. Results are expressed as the mean ear swelling±SD. Data are representative of four experiments. Journal of Investigative Dermatology 2007 127, 1430-1435DOI: (10.1038/sj.jid.5700726) Copyright © 2007 The Society for Investigative Dermatology, Inc Terms and Conditions