Resolution of endothelial activation and down-regulation of Tie2 receptor in psoriatic skin after infliximab therapy Trevor Markham, MD, Ronan Mullan,

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Resolution of endothelial activation and down-regulation of Tie2 receptor in psoriatic skin after infliximab therapy Trevor Markham, MD, Ronan Mullan, MRCP, Lucy Golden-Mason, PhD, Sarah Rogers, MD, Barry Bresnihan, MD, Oliver FitzGerald, MD, Ursula Fearon, PhD, Douglas J. Veale, MD Journal of the American Academy of Dermatology Volume 54, Issue 6, Pages 1003-1012 (June 2006) DOI: 10.1016/j.jaad.2006.01.038 Copyright © 2006 American Academy of Dermatology, Inc. Terms and Conditions

Fig 1 Representative photographs of patient with psoriasis pre- and post-infliximab therapy. A and B, Extensive psoriasis covering trunk pretreatment. C and D, After week 12, post-infliximab therapy psoriasis has completely cleared. Journal of the American Academy of Dermatology 2006 54, 1003-1012DOI: (10.1016/j.jaad.2006.01.038) Copyright © 2006 American Academy of Dermatology, Inc. Terms and Conditions

Fig 2 Clinical effects of infliximab (3-5 mg/mL) on mean Psoriasis Area and Severity Index (PASI) and body surface area (BSA) scores from week 0 to 12 after 3 infusions at baseline and 2 and 6 weeks (n = 15). Results are expressed as mean ± SEM. ∗P < .05 significantly different from baseline. Journal of the American Academy of Dermatology 2006 54, 1003-1012DOI: (10.1016/j.jaad.2006.01.038) Copyright © 2006 American Academy of Dermatology, Inc. Terms and Conditions

Fig 3 Clinical effect of infliximab therapy (3-5 mg/kg). Disease Activity Scores (DAS) and Health Assessment Questionaire (HAQ) joint scores (n = 8) pre-infliximab (week 0) and after 3 infusions of infliximab (week 12). (∗P < .05). Journal of the American Academy of Dermatology 2006 54, 1003-1012DOI: (10.1016/j.jaad.2006.01.038) Copyright © 2006 American Academy of Dermatology, Inc. Terms and Conditions

Fig 4 Quantification of angiopoietin (Ang) 1, Ang 2, tumor necrosis factor (TNF)-α, and Tie2 messenger RNA (mRNA) expression by real-time polymerase chain reaction normalized to β-actin pre- and post-infliximab therapy. Results are expressed as mean ± SEM. A, Significant reductions in mRNA expression of Ang 1 and Tie2 at 12 weeks posttreatment (n = 14) (∗P < .05). B, Gel represents Tie2 mRNA in involved (lane 1) and uninvolved (lane 2) skin pretreatment, and 2 (lane 3) and 12 (lane 4) weeks post-infliximab therapy. Journal of the American Academy of Dermatology 2006 54, 1003-1012DOI: (10.1016/j.jaad.2006.01.038) Copyright © 2006 American Academy of Dermatology, Inc. Terms and Conditions

Fig 5 Immunohistochemical staining of skin biopsy specimens. Photomicrographs of immunohistochemical staining demonstrating intense staining for CD3, platelet-endothelial cell adhesion molecule (PECAM), angiopoietin (Ang) 2, and Tie2 in involved skin at baseline (A to D). At 12 weeks posttherapy there is marked decrease in staining for CD3, PECAM, Ang 2, and Tie2 (E to H). Minimal expression of CD3 and Tie2 are demonstrated in normal skin (I and J). Original magnification ×20. Journal of the American Academy of Dermatology 2006 54, 1003-1012DOI: (10.1016/j.jaad.2006.01.038) Copyright © 2006 American Academy of Dermatology, Inc. Terms and Conditions

Fig 6 Immunohistochemical staining scores (mean ± SEM) of inflammatory markers CD3, CD4, CD68, CD31, and CD34 at baseline in clinically involved (n = 15) and uninvolved (n = 15) skin (A) and post-infliximab therapy (n = 15) at 2 and 12 weeks (C). Immunohistochemical staining scores (mean ± SEM) of growth factors angiopoietin (Ang) 2, vascular endothelial growth factor (VEGF), Tie2, and tumor necrosis factor (TNF)-α at baseline in involved and uninvolved skin (B) and post-infliximab therapy at 2 and 12 weeks (D). Immunohistochemical staining was measured using validated semiquantitative scoring method (0-4) where 0 = no positive cells and 4 > 75% positive cells. ∗P < .05 Significant difference between baseline involved and uninvolved skin. #P < .05 Significant reduction in expression of all inflammatory markers and growth factors at week 2 and 12 compared with pretreatment involved scores. Journal of the American Academy of Dermatology 2006 54, 1003-1012DOI: (10.1016/j.jaad.2006.01.038) Copyright © 2006 American Academy of Dermatology, Inc. Terms and Conditions