Comparison of a novel CXCL12/CCL5 dependent migration assay with CXCL8 secretion and CD86 expression for distinguishing sensitizers from non-sensitizers using MUTZ-3 Langerhans cells.
Comparison of a novel CXCL12/CCL5 dependent migration assay with CXCL8 secretion and CD86 expression for distinguishing sensitizers from non-sensitizers using MUTZ-3 Langerhans cells.
复制标题
使用 MUTZ-3 Langerhans 细胞对新型 CXCL12/CCL5 依赖性迁移测定与 CXCL8 分泌和 CD86 表达进行比较，以区分敏化剂和非敏化剂。
DOI：
10.1016/j.tiv.2009.10.014
复制
发表时间：
2010
期刊：
影响因子：
--
通讯作者：
S. Gibbs
中科院分区：
文献类型：
--
作者：
K. Ouwehand;S. Spiekstra;J. Reinders;R. Scheper;T. D. de Gruijl;S. Gibbs
由于接触性超敏反应的诱导是一系列细胞过程的结果，包括表皮树突细胞（朗格汉斯细胞（LC））的成熟和迁移，因此基于这些体内事件的一系列测定可能提供强大的体外可预测性模型，用于区分致敏剂和非致敏剂。因此，将读出 CD86 表达和 CXCL8 分泌变化的测定与基于 LC 体外迁移行为的新型功能测定进行比较。在所有三个测定中，均使用源自人骨髓白血病细胞系 MUTZ-3 (MUTZ-LC) 的 LC。将 MUTZ-LC 暴露于一组五种敏化剂和三种非敏化剂中，仅导致 3/5 敏化剂中 CD86 表达增加，但也导致 1/3 非敏化剂中 CD86 表达增加。相反，在暴露于所有致敏剂后，CXCL8的分泌均匀增加，但在暴露于非致敏剂后则没有。在 Transwell 迁移测定中，观察到敏化剂暴露的 MUTZ-LC 向 CXCL12 优先迁移（5/5 敏化剂），而非敏化剂暴露的 MUTZ-LC 仅向 CCL5 迁移（3/3 非敏化剂）。总之，新型 MUTZ-LC 迁移测定和 CXCL8 分泌分析在预测致敏剂和非致敏剂方面比 CD86 分析更成功，因此值得在体外测定开发领域进行进一步研究。
As the induction of contact hypersensitivity is the result of a series of cellular processes, including maturation and migration of epidermal dendritic cells (Langerhans cells (LC)), a battery of assays based on these in vivo events might provide a robust in vitro predictability model for distinguishing sensitizers from non-sensitizers. Therefore, assays with read-out for changes in CD86 expression and CXCL8 secretion were compared with a novel functional assay based on the in vitro migratory behaviour of LC. In all three assays LC derived from the human myeloid-leukaemia-cell-line MUTZ-3 (MUTZ-LC) were used. Exposure of MUTZ-LC to a panel of five sensitizers and three non-sensitizers resulted in increased CD86 expression in only 3/5 sensitizers, but also in 1/3 non-sensitizers. In contrast, CXCL8 secretion was uniformly increased after exposure to all sensitizers, but not after exposure to non-sensitizers. In a transwell migration assay, preferential migration of sensitizer-exposed MUTZ-LC towards CXCL12 was observed (5/5 sensitizers), whereas non-sensitizer-exposed MUTZ-LC only migrated towards CCL5 (3/3 non-sensitizers). In conclusion, the novel MUTZ-LC migration assay and analysis of CXCL8 secretion proved to be more successful than analysis of CD86 in predicting sensitizers from non-sensitizers and therefore warrant further investigation in the field of in vitro assay development.