Genetic Variants of the IL22 Promoter Associate to Onset of Psoriasis before Puberty and Increased IL-22 Production in T Cells  Pernilla Nikamo, Stanley Cheuk, Josefin Lysell, Charlotta Enerbäck, Kerstin Bergh, Ning Xu Landén, Liv Eidsmo, Mona Ståhle  Journal of Investigative Dermatology  Volume 134, Issue 6, Pages 1535-1541 (June 2014) DOI: 10.1038/jid.2014.5 Copyright © 2014 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 1 Allelic association to genetic variants in the IL22 promoter in psoriasis patients stratified for age at disease onset. The 3-D graph shows a case–control association study in which psoriasis patients with onset of disease at <10 years significantly associate to genetic variants in the IL22 promoter. *P<0.05, **P<0.01. Journal of Investigative Dermatology 2014 134, 1535-1541DOI: (10.1038/jid.2014.5) Copyright © 2014 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 2 Luciferase reporter assay demonstrating a functional difference between high-risk and low-risk INDEL genotypes for the IL22 promoter. (a) Schematic showing plasmid constructs used for luciferase promoter reporter assay. (b) HeLa cells transfected with plasmid constructs containing high- or low-risk IL22 promoter genotypes were either unstimulated or stimulated with PMA. Luciferase reporter assay results from one representative experiment, with six replicates, out of four independent experiments. Student’s t-test, *P<0.05. NS, not significant. Journal of Investigative Dermatology 2014 134, 1535-1541DOI: (10.1038/jid.2014.5) Copyright © 2014 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 3 The high-risk haplotype is associated with elevated IL-22 production in ex vivo stimulated CD4 T cells from psoriasis patients with onset of disease <10 years. (a) Plasma levels of IL-22 in high-risk (n=12) or low-risk patients (n=10). (b) Representative FACS plots depicting IL-22 and IL-17 in CD4 T cells after a 48 hour incubation in the presence (anti-CD3) or absence (medium) of CD3. (c, d) Dot plots depicting frequencies of (c) IL-22- and (d) IL-17A-expressing CD4 T cells. Each dot represents one individual (high-risk: black circles, n=12, low-risk: white circles, n=11). Horizontal lines equal median. (e) Cytokine concentration in supernatants collected from high-risk (n=12) or low-risk (n=11) genotypes. Mean and standard deviation is shown. Mann–Whitney analysis was performed in a–e. *P<0.05, **P<0.01. NS, not-significant. Journal of Investigative Dermatology 2014 134, 1535-1541DOI: (10.1038/jid.2014.5) Copyright © 2014 The Society for Investigative Dermatology, Inc Terms and Conditions