title: Monitoring Skin Dendritic Cells in Steady State and Inflammation by Immunofluorescence Microscopy and Flow Cytometry.
abstract: Skin dendritic cells (DC) are strategically positioned at the body's second largest epithelial border to the environment. Hence they are the first antigen presenting cells that encounter invading pathogens and environmental antigens, including contact sensitizers and carcinogens penetrating the skin. Moreover, DC have the unique ability to induce immunity or tolerance and thus take center stage in regulating innate and adaptive immune responses. Skin DC can be divided into several phenotypically and functionally distinct subtypes. The three main subsets are Langerin+epidermal Langerhans cells (LC) and Langerin+as well as Langerinnegdermal DC. In the steady state skin DC form a dense network to survey the periphery for pathogens and harmful substances breaching the cutaneous barrier. During inflammation DC become rapidly activated and start their migration to skin-draining lymph nodes where they initiate antigen-specific T cell responses. The homeostasis and mobilization of DC in the skin can be visualized by immunofluorescent staining of epidermal and dermal sheet preparations or skin sections. Here, we describe in detail how inflammation can be induced in the skin with tape stripping or FITC painting and how the skin DC network can be monitored using immunofluorescence microscopy and flow cytometry.
authors.affiliation: Institute for Molecular Medicine, University Medical Center of the Johannes Gutenberg-University, Obere Zahlbacher Straße 67, 55131, Mainz, Germany.
authors.forename: Julia L
authors.initials: JL
authors.lastname: Ober-Blöbaum
authors.affiliation: Division of Experimental Dermatology, Department of Dermatology, Venereology and Allergology, Medical University of Innsbruck, Anichstrasse 35, A-6020, Innsbruck, Austria.
authors.forename: Daniela
authors.initials: D
authors.lastname: Ortner
authors.affiliation: Department for Pediatric Urology, Hospital of the Sisters of Charity, Linz, Austria.
authors.forename: Bernhard
authors.initials: B
authors.lastname: Haid
authors.affiliation: Institute for Molecular Medicine, University Medical Center of the Johannes Gutenberg-University, Obere Zahlbacher Straße 67, 55131, Mainz, Germany.
authors.forename: Anna
authors.initials: A
authors.lastname: Brand
authors.affiliation: Division of Experimental Dermatology, Department of Dermatology, Venereology and Allergology, Medical University of Innsbruck, Anichstrasse 35, A-6020, Innsbruck, Austria.
authors.forename: Christoph
authors.initials: C
authors.lastname: Tripp
authors.affiliation: Institute for Molecular Medicine, University Medical Center of the Johannes Gutenberg-University, Obere Zahlbacher Straße 67, 55131, Mainz, Germany. bclausen@uni-mainz.de.
authors.forename: Björn E
authors.initials: BE
authors.lastname: Clausen
authors.affiliation: Division of Experimental Dermatology, Department of Dermatology, Venereology and Allergology, Medical University of Innsbruck, Anichstrasse 35, A-6020, Innsbruck, Austria. patrizia.stoitzner@i-med.ac.at.
authors.forename: Patrizia
authors.initials: P
authors.lastname: Stoitzner
mesh_terms: D000818:Animals; D015703:Antigens, CD / Q000235:genetics / Q000276:immunology; D002465:Cell Movement; D003713:Dendritic Cells / Q000145:classification / Q000276:immunology*; D020405:Dermis / Q000276:immunology* / Q000473:pathology; D004423:Ear; D004817:Epidermis / Q000276:immunology* / Q000473:pathology; D005434:Flow Cytometry / Q000379:methods*; D016650:Fluorescein-5-isothiocyanate / Q000737:chemistry; D005456:Fluorescent Dyes / Q000737:chemistry; D006204:Hair Removal / Q000295:instrumentation / Q000379:methods; D007249:Inflammation; D051379:Mice; D008810:Mice, Inbred C57BL; D008856:Microscopy, Fluorescence / Q000379:methods*; D013194:Staining and Labeling / Q000379:methods*; D053832:Surgical Tape
chemical_list: D015703:Antigens, CD; D005456:Fluorescent Dyes; D016650:Fluorescein-5-isothiocyanate
keywords: Dendritic cells; Epidermal and dermal sheets; FITC painting; Immunofluorescence microscopy; Langerhans cells; Skin cryosections; Skin inflammation; Tape stripping