The Angiogenesis Inhibitor Thrombospondin-1 Inhibits Acute Cutaneous Hypersensitivity Reactions  Paula Velasco, Rainer Huegel, Jochen Brasch, Jens M. Schröder, Michael Weichenthal, Eggert Stockfleth, Thomas Schwarz, Jack Lawler, Michael Detmar, Bernhard Lange-Asschenfeldt  Journal of Investigative Dermatology  Volume 129, Issue 8, Pages 2022-2030 (August 2009) DOI: 10.1038/jid.2008.447 Copyright © 2009 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 1 Upregulation of TSP-1 expression and its receptor CD36 in human acute contact dermatitis. Immunohistochemical stains of (a) uninflamed and (b) inflamed back skin at 72hours after the onset of a human ACD (acute allergic contact dermatitis) using an antibody directed against human TSP-1. Non-radioactive in situ hybridization (ISH) using a riboprobe specific for human TSP-1 in the uninflamed human skin (c). In situ hybridization for TSP-1 (red) combined with an immunostain for collagen IV (green) revealed TSP-1 mRNA-positive vessels (arrows) in the inflamed skin (d). Double immunofluorescence stain with antibodies directed against laminin (red) and TSP-1 (green) revealed TSP-1-labeled vessels (yellow, arrowheads) (e). Immunohistochemical stains of uninflamed (f) and inflamed skin (g) using an antibody directed against CD36. Double immunofluorescence stains of inflamed skin with antibodies directed against laminin (red) and CD36 (green) reveal a scattered staining pattern in the dermis and the epidermis (arrows) (h). White dots delineate the dermoepidermal junction. Twelve uninflamed and 15 inflamed skin samples were analyzed. Bars=200μm (a, b, and d–h), 100μm (c). Journal of Investigative Dermatology 2009 129, 2022-2030DOI: (10.1038/jid.2008.447) Copyright © 2009 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 2 Strong expression of TSP-1 protein in the inflamed murine skin. Immunohistochemical stains of uninflamed (a) and inflamed (b) skin of wild-type mice revealed TSP-1 protein expression in the epidermis. Bars=200μm. Journal of Investigative Dermatology 2009 129, 2022-2030DOI: (10.1038/jid.2008.447) Copyright © 2009 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 3 Reduced vascular leakage and vascularity in the inflamed skin of TSP-1 transgenic mice. In (a) wild-type and (b) TSP-1 transgenic mice, a mouse ear swelling test (c) was performed after the induction of CHS reactions. At least six TSP-1 transgenic and six wild-type mice per time point were used for the experiment. Ear swelling is expressed as the increase (change) over the original ear thickness in micrometers. Based on CD 31-immunostained vessels in the tissue of wild-type (d) and TSP-1 transgenic mice (e) computer-assisted morphometric image analyses were performed (f) at 24 hours after the onset of inflammation. Skin samples from at least six TSP-1 transgenic and six wild-type mice were analyzed. Data are representative of three independent experiments. Data are expressed as median and range. **P<0.01 (U-test). Bars=200μm. Journal of Investigative Dermatology 2009 129, 2022-2030DOI: (10.1038/jid.2008.447) Copyright © 2009 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 4 Enhanced inflammation and increased vascular remodeling in the skin of TSP-1-deficient mice Ear swelling of (a) wild-type and (b) TSP-1-deficient mice after the induction of CHS reactions (c). Ear swelling is expressed as the increase (change) over the original ear thickness in micrometers (c). Based on CD31-stained cryosections of the inflamed ear skin of wild-type (d) and TSP-1-deficient mice (e) morphometric image analyses were performed (f) 24 hours after the induction of a CHS reaction. Six TSP-1-deficient and six wild-type mice per time point were analyzed. Reported data are representative of three independent experiments. Data are expressed as median and range. *P<0.05; **P<0.01 (U-test). Bar=200μm. Journal of Investigative Dermatology 2009 129, 2022-2030DOI: (10.1038/jid.2008.447) Copyright © 2009 The Society for Investigative Dermatology, Inc Terms and Conditions