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Mouse Models for Atopic Dermatitis
Keiko Sakamoto1,2, Keisuke Nagao1
1Cutaneous Leukocyte Biology Section, Dermatology Branch, National Institute of Arthritis and Musculoskeletal and Skin Diseases, National Institutes of Health, Bethesda, Maryland.
Abstract:
Atopic dermatitis (AD) is a multifactorial disease with underlying barrier disruption and altered microbial flora, resulting in dry skin and eczematous inflammation with persistent pruritis. Mouse models have been heavily used to investigate AD pathophysiology. Among various AD mouse models, AD-like inflammation induced by topical calcipotriol, a vitamin D3 analog referred to as MC903 in experimental settings, is a versatile model that can be applied to any strain of mice, which can be used for immunologic and morphologic studies. Herein, we provide basic protocols for the topical application of MC903 and approaches to assess phenotypes. After inducing AD-like inflammation, the skin is harvested for flow cytometry analysis, as well as for histologic and immunofluorescence microscopy analyses. The combination of these approaches enables accurate characterization of the degree of inflammation, type of inflammatory infiltrate, and localization of immune infiltrates. Published 2023. This article is a U.S. Government work and is in the public domain in the USA. Basic Protocol 1: Application of MC903 and gross phenotype assessment Basic Protocol 2: Processing skin for flow cytometry analysis Support Protocol: Skin immune cell surface staining and flow cytometry analysis Basic Protocol 3: Harvesting skin for histologic analysis Basic Protocol 4: Immunofluorescence staining to identify immune cell infiltrates.
Insights
This study details a versatile mouse model for atopic dermatitis (AD) using topical MC903 (calcipotriol) to induce inflammation. Protocols are provided for assessing skin phenotypes, immune cell infiltration, and inflammation using flow cytometry and microscopy.
Area of Science:
- Immunology
- Dermatology
- Animal Models
Background:
- Atopic dermatitis (AD) is a complex skin condition characterized by barrier dysfunction, inflammation, and itching.
- Mouse models are crucial for understanding AD pathophysiology.
- The MC903 (calcipotriol) topical application model offers a versatile method for inducing AD-like inflammation in various mouse strains.
Purpose of the Study:
- To provide standardized protocols for inducing and assessing AD-like inflammation in mice using topical MC903.
- To detail methods for evaluating skin phenotypes, immune cell infiltration, and inflammatory markers.
Main Methods:
- Topical application of MC903 to induce AD-like skin inflammation.
- Gross phenotype assessment of skin lesions.
- Skin processing for flow cytometry to analyze immune cell populations.
- Histologic and immunofluorescence microscopy for detailed analysis of inflammation and immune cell localization.
Main Results:
- The MC903 model allows for reproducible induction of AD-like inflammation.
- Combined analyses (flow cytometry, histology, immunofluorescence) enable comprehensive characterization of the inflammatory response.
- Accurate assessment of inflammation degree, inflammatory infiltrate type, and immune cell localization is achievable.
Conclusions:
- The MC903-induced mouse model is a valuable tool for studying AD pathogenesis.
- Standardized protocols facilitate robust immunologic and morphologic investigations of AD.
- This model aids in understanding the complex immune mechanisms underlying atopic dermatitis.
