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Human keratinocytes and monocytes co-culture cell system: An important contribution for the study of moderate and weak sensitizers.
Toxicology in Vitro ( IF 3.2 ) Pub Date : 2020-07-03 , DOI: 10.1016/j.tiv.2020.104929
Valentina Galbiati 1 , Ambra Maddalon 1 , Martina Iulini 2 , Marina Marinovich 1 , Emanuela Corsini 2
Affiliation  

In vitro approaches to address key events in chemical-induced skin sensitization have been developed, but there is still uncertain how they will be useful to predict the potency for an effective risk assessment. Keratinocytes (KCs) play a key role in all phases of skin sensitization. Dendritic cells (DCs) activation and maturation require the binding of cytokines produced by KC as a result of initial chemical exposure. We previously identified interleukin-18 (IL-18) as useful marker for determination of skin sensitization potential of chemicals. The aim of this paper was to mimic the interaction between KCs and DCs using a co-culture of NCTC 2544 and THP-1 cells. Three selected contact allergens of different potency (Bandrowski's base, diethyl maleate, and imidazolidinyl urea) were tested in time-course experiments (24, 48 and 72 h). Cell surface markers expression (CD80, CD86, and HLA-DR) was determined by flow cytometry analysis while IL-18 production was evaluated with specific sandwich ELISA. Results obtained from this simple in vitro co-culture system show the possibility to study the contribution of KCs in DCs activation through the analysis of HLA-DR expression. Results obtained demonstrate the ability of the KCs to favor the full maturation of the DCs in the presence of moderate and weak allergens, while the extreme allergen induced a complete maturation of DC alone without the need of KCs.



中文翻译:

人角质形成细胞和单核细胞共培养细胞系统:对中敏和弱敏剂研究的重要贡献。

体外已经开发出用于解决化学诱导的皮肤致敏中的关键事件的方法,但是仍然不确定它们将如何用于预测有效风险评估的效力。角质形成细胞(KCs)在皮肤敏化的所有阶段均起关键作用。树突状细胞(DCs)的激活和成熟需要结合KC产生的细胞因子,因为最初的化学暴露会导致这种细胞因子的结合。我们先前将白介素18(IL-18)鉴定为确定化学物质对皮肤致敏潜力的有用标记。本文的目的是使用NCTC 2544和THP-1细胞的共培养物来模拟KC和DC之间的相互作用。在时程实验(24、48和72小时)中测试了三种选择的不同效价的接触过敏原(Bandrowski碱,马来酸二乙酯和咪唑烷基脲)。通过流式细胞术分析确定细胞表面标志物的表达(CD80,CD86和HLA-DR),同时通过特异性夹心ELISA评估IL-18的产生。从这个简单的结果体外共培养系统显示了通过分析HLA-DR表达来研究KC在DC激活中的作用的可能性。获得的结果表明,在中等和较弱的变应原存在的情况下,KC促进DC完全成熟的能力,而极端变应原可单独诱导DC完全成熟,而无需KC。

更新日期:2020-07-10
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