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Dorothea Dijkstra, Holger Stark, Paul L. Chazot, Fiona C

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Presentation on theme: "Dorothea Dijkstra, Holger Stark, Paul L. Chazot, Fiona C"— Presentation transcript:

1 Human Inflammatory Dendritic Epidermal Cells Express a Functional Histamine H4 Receptor 
Dorothea Dijkstra, Holger Stark, Paul L. Chazot, Fiona C. Shenton, Rob Leurs, Thomas Werfel, Ralf Gutzmer  Journal of Investigative Dermatology  Volume 128, Issue 7, Pages (July 2008) DOI: /sj.jid Copyright © 2008 The Society for Investigative Dermatology, Inc Terms and Conditions

2 Figure 1 Expression of the H4R by IDECs in skin sections. Skin sections (1μm) of formalin-fixed and paraffin-embedded lesional AD skin biopsies were stained with either FITC–CD36 or FITC–CD206 (to identify IDECs) and counterstained for the expression of the H4R (Texas Red). One representative experiment of five is shown. Bar=20μm. Journal of Investigative Dermatology  , DOI: ( /sj.jid ) Copyright © 2008 The Society for Investigative Dermatology, Inc Terms and Conditions

3 Figure 2 Expression of the H4R by IDECs in epidermal cell suspensions performed on freshly obtained shave biopsies from lesional AD skin. IDECs were stained for CD206 (FITC) and H4R (peridinin–chlorophyll–protein complex), CD206-positive cells were gated and analyzed for H4R expression. (a) One representative experiment is shown. GM, geometric mean of fluorescence. (b) Nine experiments are summarized. Statistical analysis was performed by Wilcoxon's signed-rank test; **P<0.005. Journal of Investigative Dermatology  , DOI: ( /sj.jid ) Copyright © 2008 The Society for Investigative Dermatology, Inc Terms and Conditions

4 Figure 3 Expression and regulation of the H4R on Mo-IDECs. (a) Staining of H4R (thick line), isotype (gray filled thin line), and blocked H4R antibody (thin line). (b) Regulation of H4R expression by IFN-γ (20ngml−1), IL-4 (20ngml−1), TNF-α (0.5ngml−1), poly I:C (TLR3 ligand, 2μgml−1), or combinations of these (mean±SEM of five independent experiments is shown). Statistical analysis was performed by ANOVA; **P< (c) Upregulation by IFN-γ, IFN-γ+TNF-α and IFN-γ+TNF-α and IFN-γ+TNF-α+poly I:C (thick lines) as compared with non-stimulated cells (thin lines) in one representative experiment. Journal of Investigative Dermatology  , DOI: ( /sj.jid ) Copyright © 2008 The Society for Investigative Dermatology, Inc Terms and Conditions

5 Figure 4 Histamine downregulates CCL2 protein via H4R. Mo-IDECs were stimulated with HA or agonists of H1R (β-histine; β his), H2R (dimaprit; Dim), H3R (methimepip; MP), and H4R (Clob and 4-MH), for 48hours at 10μM or at the indicated concentrations. JNJ (JNJ) was used as an H4R-neutral antagonist. CCL2 was measured by ELISA, normalized to non-stimulated cells: 3,261±1,902ngml−1 (mean±SEM). (a) HA downregulated CCL2 solely via H4R. (b) Dose–response curves of CCL2 downregulation by HA, Clob, and 4-MH. Mean±SEM of five different experiments are shown. Statistical analysis was performed by ANOVA; *P<0.05, **P<0.005. Journal of Investigative Dermatology  , DOI: ( /sj.jid ) Copyright © 2008 The Society for Investigative Dermatology, Inc Terms and Conditions

6 Figure 5 H4R agonist stimulation downregulates chemotaxis induced by supernatants from Mo-IDECs. Supernatants from Mo-IDECs stimulated for 48hours with various H4R ligands were used as chemoattractants. Values normalized to chemotaxis induced by supernatant from non-stimulated Mo-IDECs (NS). HA, Clob, and 4-MH (10μM), CCL2 neutralizing antibody (a-CCL2). Mean±SEM of four experiments are shown. Statistical analysis was performed by ANOVA; *P<0.05, **P<0.005, ***P< Journal of Investigative Dermatology  , DOI: ( /sj.jid ) Copyright © 2008 The Society for Investigative Dermatology, Inc Terms and Conditions

7 Figure 6 Histamine downregulates IL-12p40 via H4R. Mo-IDECs were incubated with the same ligands as described in Figure 4 for 48hours. After 24hours of incubation poly I:C (2μgml−1) was added to the culture as second stimulus. Values are normalized to IL-12p40 levels produced by IDEC stimulated only with poly I:C (2,950±2,340pgml−1, mean±SEM). Statistical analysis was performed by ANOVA; **P<0.005. Journal of Investigative Dermatology  , DOI: ( /sj.jid ) Copyright © 2008 The Society for Investigative Dermatology, Inc Terms and Conditions


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