UV-B radiation induces the expression of antimicrobial peptides in human keratinocytes in vitro and in vivo  Regine Gläser, MD, Fatemeh Navid, PhD, Winfried Schuller, PhD, Christian Jantschitsch, MD, Jürgen Harder, PhD, Jens M. Schröder, PhD, Agatha Schwarz, PhD, Thomas Schwarz, MD  Journal of Allergy and Clinical Immunology  Volume 123, Issue 5, Pages 1117-1123 (May 2009) DOI: 10.1016/j.jaci.2009.01.043 Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 1 UV radiation induces antimicrobial peptides at the mRNA level in normal human keratinocytes. Normal human keratinocytes were UV-irradiated (0, 150, 350 J/m2). RNA was extracted 24 hours later and reverse-transcribed, and quantitative real-time PCR was performed by using primers specific for HBD-2, HBD-3, RNase 7, and psoriasin. The 2−ΔΔCt values are the results of 1 representative experiment from 3 independent experiments. Journal of Allergy and Clinical Immunology 2009 123, 1117-1123DOI: (10.1016/j.jaci.2009.01.043) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 2 UV radiation induces antimicrobial peptides at the mRNA level in normal human keratinocytes. Normal human keratinocytes were UV-irradiated (350 J/m2). RNA was extracted 4, 8, 24, or 48 hours later and reverse-transcribed, and quantitative real-time PCR was performed by using primers specific for HBD-2, HBD-3, RNase 7, and psoriasin. Unirradiated cells served as controls (Co). The 2−ΔΔCt values are the results of 1 representative experiment from 3 independent experiments. Journal of Allergy and Clinical Immunology 2009 123, 1117-1123DOI: (10.1016/j.jaci.2009.01.043) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 3 UV radiation induces antimicrobial peptides at the protein level in normal human keratinocytes. A, Normal human keratinocytes were UV-irradiated (0, 150, 350 J/m2). Twenty-four hours later, cells were harvested, and intracellular staining was performed by using antibodies against HBD-2, HBD-3, RNase 7, and psoriasin followed by FACS analysis. Specific fluorescence (X-Mean) was calculated using isotype controls with X Mean values set as 1. Data show 1 representative of seven independent experiments. B, Keratinocytes were exposed to 350 J/m2 UV radiation or left unirradiated (Co). Twenty-four hours later, cells were fixed with methanol and incubated with antibodies against HBD-2, HBD-3, RNase 7, and psoriasin. Bound antibodies were visualized by Alexa488-labeled secondary antibodies. Analysis was performed with a fluorescence microscope. Bar: 200 μm. Journal of Allergy and Clinical Immunology 2009 123, 1117-1123DOI: (10.1016/j.jaci.2009.01.043) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 4 UV radiation induces antimicrobial peptides in vivo. Six healthy volunteers were irradiated on 3 consecutive days with 1 MED UV-B on their buttocks. Punch biopsies were taken from these areas before and 1 day and 6 days after UV exposure Biopsies of unirradiated skin serves as controls (Co). Immunohistochemical analysis was performed by using antibodies directed against HBD-2, HBD-3, RNase 7, and psoriasin. Samples from 1 representative person are shown. Bar: 50 μm. Journal of Allergy and Clinical Immunology 2009 123, 1117-1123DOI: (10.1016/j.jaci.2009.01.043) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 5 UV radiation induces antimicrobial peptides in skin explants. Skin explants derived from plastic surgery samples were UV-B–irradiated (250 J/m2). Twenty-four hours later, punch biopsies were taken and immunohistochemical analysis was performed by using antibodies directed against HBD-2, HBD-3, RNase 7, and psoriasin. Unirradiated samples served as controls (Co). Samples of 1 representative experiment out of 6 are shown. Bar: 100 μm. Journal of Allergy and Clinical Immunology 2009 123, 1117-1123DOI: (10.1016/j.jaci.2009.01.043) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions