Institute of Environmental Medicine, Karolinska Institute, Stockholm, Sweden.
J Immunotoxicol. 2021 Dec;18(1):74-84. doi: 10.1080/1547691X.2021.1905754.
Sensitization to a contact allergen brings with it a lifelong risk to develop allergic contact dermatitis. Inflammation is an important part of the skin sensitizing mechanism, and understanding how different haptens stimulate the immune system, as well as the role played by different cell types present in skin, may be helpful for developing optimized models for risk assessment of new chemicals or mixtures. The aim of this study was to compare the cytokine profile following exposure of cells representing keratinocytes (HaCaT), monocytes (THP-1) and a co-culture of these cells to three clinically important skin sensitizers: cobalt (II) chloride (CoCl), methylisothiazolinone (MI) and p-phenylenediamine (PPD). Secretion of ten pro-inflammatory cytokines was measured using multiplexing. The results showed that the cytokine response differed substantially between the three cell assays. CoCl caused an increase of IL-8 in HaCaT cells, while the induction of also IL-13 and IL-1β was observed in THP-1 cells and co-cultures. MI induced six cytokines in HaCaT cells but only IL-1β in the THP-1 cells and four cytokines in the co-culture. Interestingly, the IL-1β response was massive in the co-culture. PPD caused release of IL-1β in all three models as well as IL-8 in the co-culture. Control experiments with two non-sensitizers and irritants (lactic acid and sodium dodecyl sulfate) showed no effect on IL-8 or IL-1β in the co-culture. Taken together, results from this exploratory analysis show unique cytokine profiles dependent on the type of hapten and cell model. Importantly, all three haptens triggered secretion of IL-1β and IL-8 in a co-culture of HaCaT cells and THP-1 cells, representing the most robust test system.
变应原致敏会带来终生罹患过敏性接触性皮炎的风险。炎症是皮肤致敏机制的重要组成部分，了解不同半抗原如何刺激免疫系统，以及皮肤中不同类型细胞所起的作用，可能有助于开发优化的新化学物质或混合物风险评估模型。本研究的目的是比较三种临床上重要的皮肤致敏剂：氯化钴（CoCl）、甲基异噻唑啉酮（MI）和对苯二胺（PPD）作用于代表角质形成细胞（HaCaT）、单核细胞（THP-1）和这些细胞共培养物的细胞后细胞因子谱。使用多重分析测量了十种促炎细胞因子的分泌。结果表明，三种细胞测定法的细胞因子反应有很大差异。CoCl 引起 HaCaT 细胞中 IL-8 的增加，而在 THP-1 细胞和共培养物中观察到 IL-13 和 IL-1β 的诱导。MI 在 HaCaT 细胞中诱导六种细胞因子，但在 THP-1 细胞中仅诱导 IL-1β，在共培养物中诱导四种细胞因子。有趣的是，共培养物中的 IL-1β 反应非常强烈。PPD 在所有三种模型中均引起 IL-1β 和共培养物中 IL-8 的释放。两种非致敏剂和刺激性物质（乳酸和十二烷基硫酸钠）的对照实验显示，共培养物中 IL-8 或 IL-1β 无变化。总的来说，这项探索性分析的结果表明，细胞因子谱取决于半抗原和细胞模型的类型。重要的是，所有三种半抗原均在 HaCaT 细胞和 THP-1 细胞的共培养物中触发了 IL-1β 和 IL-8 的分泌，这是最具活力的测试系统。
