A Human Endogenous Retrovirus K dUTPase Triggers a TH1, TH17 Cytokine Response: Does It Have a Role in Psoriasis? Maria-Eugenia Ariza, Marshall V. Williams

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A Human Endogenous Retrovirus K dUTPase Triggers a TH1, TH17 Cytokine Response: Does It Have a Role in Psoriasis? Maria-Eugenia Ariza, Marshall V. Williams Journal of Investigative Dermatology Volume 131, Issue 12, Pages 2419-2427 (December 2011) DOI: 10.1038/jid.2011.217 Copyright © 2011 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 1 Deoxyuridine triphosphate nucleotidohydrolase (dUTPase) protein blast alignment. Human endogenous retrovirus K (HERV-K) consensus, as described by Harris et al. (1997), low-risk haplotype and high-risk haplotype, as described by Foerster et al. (2005), HERV-K dUTPase recombinant proteins reflecting wild-type, low-risk (N158K and K155R/N158K) and high-risk (N158R and K155G/N158R) haplotypes, as well as various deletion mutations (Δ162–171; Δ155–171, Δ142–171) used in this study, and the human dUTPase (HudUT). Boxes D-1–D-5 represent the conserved domains of the dUTPase protein family. Journal of Investigative Dermatology 2011 131, 2419-2427DOI: (10.1038/jid.2011.217) Copyright © 2011 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 2 SDS-PAGE of purified human endogenous retrovirus K (HERV-K) deoxyuridine triphosphate nucleotidohydrolase (dUTPase). SDS-PAGE was performed as described in Materials and Methods using a 10% gel. Approximately 25μg of purified protein was added to each lane. The following electrophoresis gels were stained with Lab Safe Gel Blue according to the manufacturer's instructions. Lane 1: wild-type (WT) HERV-K dUTPase; lane 2: N158K; lane 3: K155R/N158K; lane 4: N158R; lane 5: K155G/N158R; lane 6: Δ142–171; lane 7: Δ155–171; lane 8: Δ162–171. Journal of Investigative Dermatology 2011 131, 2419-2427DOI: (10.1038/jid.2011.217) Copyright © 2011 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 3 Human endogenous retrovirus K (HERV-K) deoxyuridine triphosphate nucleotidohydrolase (dUTPase) modulates innate immunity. HERV-K dUTPase proteins activate NF-κB through TLR2 in HEK293TLR2-expressing cells, as determined by luciferase assay. HEK293TLR2 and wild-type (WT) HEK293 cells were transiently transfected with NF-κB luciferase reporter plasmid, as described previously (Ariza et al., 2009). After 24–36hours, cells were treated with various HERV-K dUTPase proteins (10μgml−1), human dUTPase control protein (10μgml−1), and zymosan (10μgml−1, data not shown), or left untreated for 8hours, and luciferase reporter gene activity was measured. Data are expressed as the mean fold induction±SD relative to control levels. Values represent the average of three independent experiments. HEK293, human embryonic kidney 293; TLR2, Toll-like receptor 2. Journal of Investigative Dermatology 2011 131, 2419-2427DOI: (10.1038/jid.2011.217) Copyright © 2011 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 4 Human endogenous retrovirus K (HERV-K) deoxyuridine triphosphate nucleotidohydrolase (dUTPase) proteins stimulate the secretion of TH1, TH17 cytokines in human dendritic/Langerhans-like cells (hDCs). HERV-K dUTPase proteins differentially induce the secretion of TH1, TH17 cytokines in hDCs following a 24-hour treatment, as determined by proteome array. (a) IL-23 and IL12p40; (b) tumor necrosis factor (TNF)-α and IL-1β; (c) IL-17 and IFN-γ; (d) CCL20 and RANTES; (e) IL-8 and IL-6; (f) IL-10 and transforming growth factor (TGF)-α. Cytokine/chemokine concentrations represent the average±SD (pgml−1) from two independent experiments (n=4). *P<0.05. Journal of Investigative Dermatology 2011 131, 2419-2427DOI: (10.1038/jid.2011.217) Copyright © 2011 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 5 Effect of human endogenous retrovirus K (HERV-K) deoxyuridine triphosphate nucleotidohydrolase (dUTPase) proteins on TH1, TH17 cytokine secretion in human primary keratinocytes. HERV-K dUTPase proteins differentially induce the secretion of chemokines and proinflammatory cytokines in primary keratinocytes following a 24-hour treatment, as determined by proteome array. (a) IL-8 and CCL20; (b) transforming growth factor (TGF)-α and RANTES; (c) IL-23 and IL-12p40; (d) IL-1β and IL-17. Cytokine/chemokine concentrations represent the average±SD (pgml−1) from two independent experiments (n=4). *P<0.05. Journal of Investigative Dermatology 2011 131, 2419-2427DOI: (10.1038/jid.2011.217) Copyright © 2011 The Society for Investigative Dermatology, Inc Terms and Conditions