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 Langerinneg dermal 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.