Department of Microbiology, Biomedicine Discovery Institute, Monash University, Clayton, Victoria, Australia.
Department of Physiology, Biomedicine Discovery Institute, Monash University, Clayton, Victoria, Australia.
Curr Protoc. 2023 Mar;3(3):e695. doi: 10.1002/cpz1.695.
Atopic dermatitis (AD) is a chronic, relapsing, and extremely pruritic inflammatory skin disease with a particular impact on children. AD pathogenesis is not yet fully understood, and there is no curative treatment for this disease. Therefore, several genetically or chemically-induced AD mouse models have been developed. These preclinical mouse models are an indispensable research tool for studying AD pathogenesis and evaluating the efficacy of new candidate AD therapeutics. A commonly used mouse model of AD has been developed using the topical application of a low-calcemic analog of vitamin D3, MC903, to induce AD-like inflammatory phenotypes that closely resemble human AD. Moreover, this model shows a minimal effect on systemic calcium metabolism that is observed in the vitamin D3-induced AD model. Thus, an expanding number of studies use the MC903-induced AD model to interrogate AD pathobiology in vivo and to test new candidate small molecule and monoclonal antibody therapies. This protocol describes in detail functional measurements including the measurement of skin thickness, which is a surrogate marker for ear skin inflammation, as well as itch assessment, histological evaluation to assess the structural changes associated with AD skin inflammation, and preparation of single-cell suspensions from ear skin and draining lymph nodes for the assessment of inflammatory leukocyte subset infiltration in these tissues using flow cytometry. © 2023 The Authors. Current Protocols published by Wiley Periodicals LLC. Basic Protocol: Topical application of MC903 induces AD-like skin inflammation Support Protocol 1: Measurement of ear skin thickness Support Protocol 2: Itch assessment Support Protocol 3: Dissection of ear skin and ear draining lymph nodes Support Protocol 4: Histological evaluation and quantification Support Protocol 5: Preparation of single-cell suspension from ear skin and draining lymph nodes for the assessment of inflammatory immune cell infiltration using flow cytometry.
特应性皮炎（AD）是一种慢性、复发性且极度瘙痒的炎症性皮肤病，尤其对儿童影响较大。AD 的发病机制尚未完全阐明，且目前尚无针对该病的治愈性疗法。因此，人们开发了几种基因或化学诱导的 AD 小鼠模型。这些临床前小鼠模型是研究 AD 发病机制和评估新型候选 AD 治疗药物疗效的不可或缺的研究工具。一种常用的 AD 小鼠模型是通过局部应用低钙维生素 D3 类似物 MC903 来诱导 AD 样炎症表型，这种表型与人类 AD 非常相似。此外，与维生素 D3 诱导的 AD 模型相比，该模型对全身钙代谢的影响较小。因此，越来越多的研究使用 MC903 诱导的 AD 模型在体内研究 AD 的病理生物学，并测试新型候选小分子和单克隆抗体疗法。本方案详细描述了功能测量，包括测量作为耳部皮肤炎症替代标志物的皮肤厚度，以及瘙痒评估、组织学评估以评估与 AD 皮肤炎症相关的结构变化，以及从耳部皮肤和引流淋巴结制备单细胞悬液，用于使用流式细胞术评估这些组织中炎症性白细胞亚群的浸润。© 2023 作者。 Wiley Periodicals LLC 出版的《当代协议》。 基本方案：MC903 的局部应用可诱导 AD 样皮肤炎症 支持方案 1：耳部皮肤厚度的测量 支持方案 2：瘙痒评估 支持方案 3：耳部皮肤和耳部引流淋巴结的解剖 支持方案 4：组织学评估和定量 支持方案 5：从耳部皮肤和引流淋巴结制备单细胞悬液，用于使用流式细胞术评估炎症性免疫细胞浸润。
