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Sensitization via Healthy Skin Programs Th2 Responses in Individuals with Atopic Dermatitis  Louise Newell, Marta E. Polak, Jay Perera, Charlotte Owen,

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Presentation on theme: "Sensitization via Healthy Skin Programs Th2 Responses in Individuals with Atopic Dermatitis  Louise Newell, Marta E. Polak, Jay Perera, Charlotte Owen,"— Presentation transcript:

1 Sensitization via Healthy Skin Programs Th2 Responses in Individuals with Atopic Dermatitis 
Louise Newell, Marta E. Polak, Jay Perera, Charlotte Owen, Peter Boyd, Christopher Pickard, Peter H. Howarth, Eugene Healy, John W. Holloway, Peter S. Friedmann, Michael R. Ardern-Jones  Journal of Investigative Dermatology  Volume 133, Issue 10, Pages (October 2013) DOI: /jid Copyright © 2013 The Society for Investigative Dermatology, Inc Terms and Conditions

2 Figure 1 2,4-Dinitrochlorobenzene (DNCB) penetration through the epidermis. (a, b) DNCB (20μgcm−2) was applied to the epidermis of skin biopsies from controls, (c, d) from individuals with atopic dermatitis without filaggrin mutation (ADWT), or (e, f) from filaggrin mutation heterozygote (ADFM) individuals. DNCB penetration was analyzed by immunohistochemistry with (a–f) anti-DNCB or (g) representative antibody control (green). Penetration of DNCB was assessed by measurement of the relative fluorescence of anti-DNCB (12 independent readings) from the base of the stratum corneum vs. the basal layer, as indicated by white dotted lines, (h) using image analysis software. Error bars represent mean±SEM. NS, no statistical significance. Scale bar=10μm. Journal of Investigative Dermatology  , DOI: ( /jid ) Copyright © 2013 The Society for Investigative Dermatology, Inc Terms and Conditions

3 Figure 2 Circulating lymphocytes in atopic dermatitis (AD) show a normal Th1/2 ratio. (a, b) Sixteen-hour IFN-γ ELISpot and (c, d) IL-4 ELISpot assay of ex vivo peripheral blood mononuclear cell (PBMC) activation with either phytohemagglutinin (PHA) (a, c) or Staphylococcal enterotoxin B (SEB) (b, d) in control individuals (gray lines) or in individuals with AD (black lines). (e) Ex vivo PBMC IL-4 ELISpot stimulated with house dust mite (HDM) extract. AD, n=18; Control, n=10. *P<0.05. SFU, spot-forming units. Journal of Investigative Dermatology  , DOI: ( /jid ) Copyright © 2013 The Society for Investigative Dermatology, Inc Terms and Conditions

4 Figure 3 Atopic dermatitis (AD) subjects have impaired allergic contact dermatitis responses to 2,4-dinitrochlorobenzene (DNCB). Clinical response to epicutaneous challenge with DNCB 1 month following sensitization through the skin in AD (black lines) and controls (gray lines). Change in (a) skinfold thickness and (b) erythema index. (c) Area under the curve (AUC) for change in skinfold thickness and (d) erythema index for DNCB-elicitation reactions in controls or in individuals with AD with (ADFM) or without filaggrin mutations (ADWT). NS, no statistical significance. Journal of Investigative Dermatology  , DOI: ( /jid ) Copyright © 2013 The Society for Investigative Dermatology, Inc Terms and Conditions

5 Figure 4 2,4-Dinitrochlorobenzene (DNCB)-specific Th1 and Th2 cells. (a, d) DNCB-specific responses detected in peripheral blood mononuclear cells (PBMCs) (ex vivo; gray bars) or after short-term culture with DNCB (line; black bars) by (a) IFN-γ or (d) IL-4 ELISpot from a DNCB-sensitized atopic individual. (b, c, e, f) Flow cytometric analysis of IFN-γ or IL-4 production by (c, f) activated DNCB-expanded T-cell lines or (b, e) unstimulated control PBMCs. (a, c) Low-frequency Th1 and (d, f) Th2 cells can be detected ex vivo, but higher frequency responses are detected after short-term culture expansion. Population frequency (%) as indicated in appropriate quartiles. Representative examples are shown. SFU, spot-forming units. Journal of Investigative Dermatology  , DOI: ( /jid ) Copyright © 2013 The Society for Investigative Dermatology, Inc Terms and Conditions

6 Figure 5 Sensitization through the skin in atopic dermatitis (AD) shows unique skewing toward antigen-specific Th2 responses. ELISpot assay of 2,4-dinitrochlorobenzene (DNCB)-specific (a) IL-4 or (b) IFN-γ production by peripheral blood mononuclear cells (PBMCs) from non-sensitized controls (Non-sensitized horizontal striped bars), DNCB-sensitized controls without AD (Non-AD, black bars) (Non-AD), or DNCB-sensitized individuals with AD (AD, gray bars) 4 weeks after cutaneous DNCB sensitization AD. (c) DNCB-specific IL-4 from DNCB-sensitized controls (Non-AD), individuals without AD (ADWT) or with filaggrin mutations (ADFM). AD, n=16 (Supplementary Table S1 online); controls, n=10. Error bars represent mean±SEM. Long-lived DNCB-specific Th2 memory cells in AD shown by ELISpot assay of DNCB-specific (d) IL-4 or (e) IFN-γ production by PBMCs from controls or individuals with AD 3 months after rechallenge with DNCB (4 months after sensitization). AD, n=10; sensitized controls, n=10; non-sensitized controls, n=6. Error bars represent mean±SEM. NS, no statistical significance; SFU, spot-forming units. Journal of Investigative Dermatology  , DOI: ( /jid ) Copyright © 2013 The Society for Investigative Dermatology, Inc Terms and Conditions

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