Kosten Ilona J, Spiekstra Sander W, de Gruijl Tanja D, Gibbs Susan
Department of Dermatology, VU University Medical Center, Amsterdam, The Netherlands.
Department of Dermatology Medical Oncology, VU University Medical Center, Amsterdam, The Netherlands.
Toxicol Appl Pharmacol. 2015 Aug 15;287(1):35-42. doi: 10.1016/j.taap.2015.05.017. Epub 2015 May 29.
After allergen or irritant exposure, Langerhans cells (LC) undergo phenotypic changes and exit the epidermis. In this study we describe the unique ability of MUTZ-3 derived Langerhans cells (MUTZ-LC) to display similar phenotypic plasticity as their primary counterparts when incorporated into a physiologically relevant full-thickness skin equivalent model (SE-LC). We describe differences and similarities in the mechanisms regulating LC migration and plasticity upon allergen or irritant exposure. The skin equivalent consisted of a reconstructed epidermis containing primary differentiated keratinocytes and CD1a(+) MUTZ-LC on a primary fibroblast-populated dermis. Skin equivalents were exposed to a panel of allergens and irritants. Topical exposure to sub-toxic concentrations of allergens (nickel sulfate, resorcinol, cinnamaldehyde) and irritants (Triton X-100, SDS, Tween 80) resulted in LC migration out of the epidermis and into the dermis. Neutralizing antibody to CXCL12 blocked allergen-induced migration, whereas anti-CCL5 blocked irritant-induced migration. In contrast to allergen exposure, irritant exposure resulted in cells within the dermis becoming CD1a(-)/CD14(+)/CD68(+) which is characteristic of a phenotypic switch of MUTZ-LC to a macrophage-like cell in the dermis. This phenotypic switch was blocked with anti-IL-10. Mechanisms previously identified as being involved in LC activation and migration in native human skin could thus be reproduced in the in vitro constructed skin equivalent model containing functional LC. This model therefore provides a unique and relevant research tool to study human LC biology in situ under controlled in vitro conditions, and will provide a powerful tool for hazard identification, testing novel therapeutics and identifying new drug targets.
在接触过敏原或刺激物后，朗格汉斯细胞（LC）会发生表型变化并离开表皮。在本研究中，我们描述了源自MUTZ-3的朗格汉斯细胞（MUTZ-LC）在整合到生理相关的全层皮肤等效模型（SE-LC）中时，具有与原代朗格汉斯细胞相似的表型可塑性的独特能力。我们描述了在过敏原或刺激物暴露后调节LC迁移和可塑性的机制的异同。皮肤等效物由重建的表皮组成，其中包含原代分化的角质形成细胞和在原代成纤维细胞填充的真皮上的CD1a(+) MUTZ-LC。将皮肤等效物暴露于一系列过敏原和刺激物中。局部暴露于亚毒性浓度的过敏原（硫酸镍、间苯二酚、肉桂醛）和刺激物（曲拉通X-100、十二烷基硫酸钠、吐温80）导致LC从表皮迁移到真皮。抗CXCL12中和抗体可阻断过敏原诱导的迁移，而抗CCL5可阻断刺激物诱导的迁移。与过敏原暴露不同，刺激物暴露导致真皮内的细胞变为CD1a(-)/CDl4(+)/CD68(+)，这是MUTZ-LC在真皮中向巨噬细胞样细胞表型转换的特征。这种表型转换被抗IL-10阻断。因此，先前确定参与天然人皮肤中LC激活和迁移的机制可以在包含功能性LC的体外构建的皮肤等效模型中重现。因此，该模型提供了一种独特且相关的研究工具，用于在可控的体外条件下原位研究人类LC生物学，并将为危害识别、测试新型治疗方法和识别新的药物靶点提供强大工具。
