We have integrated dermal dendritic cell surrogates originally generated from the cell line THP-1 as central mediators of the immune reaction in a human full-thickness skin model. Accordingly, sensitizer treatment of THP-1-derived CD14-, CD11c+ immature dendritic cells (iDCs) resulted in the phosphorylation of p38 MAPK in the presence of 1-chloro-2,4-dinitrobenzene (DNCB) (2.6-fold) as well as in degradation of the inhibitor protein kappa B alpha (IκBα) upon incubation with NiSO4 (1.6-fold). Furthermore, NiSO4 led to an increase in mRNA levels of IL-6 (2.4-fold), TNF-α (2-fold) and of IL-8 (15-fold). These results were confirmed on the protein level, with even stronger effects on cytokine release in the presence of NiSO4: Cytokine secretion was significantly increased for IL-8 (147-fold), IL-6 (11.8-fold) and IL-1β (28.8-fold). Notably, DNCB treatment revealed an increase for IL-8 (28.6-fold) and IL-1β (5.6-fold). Importantly, NiSO4 treatment of isolated iDCs as well as of iDCs integrated as dermal dendritic cell surrogates into our full-thickness skin model (SM) induced the upregulation of the adhesion molecule clusters of differentiation (CD)54 (iDCs: 1.2-fold; SM: 1.3-fold) and the co-stimulatory molecule and DC maturation marker CD86 (iDCs ~1.4-fold; SM:~1.5-fold) surface marker expression. Noteworthy, the expression of CD54 and CD86 could be suppressed by dexamethasone treatment on isolated iDCs (CD54: 1.3-fold; CD86: 2.1-fold) as well as on the tissue-integrated iDCs (CD54: 1.4-fold; CD86: 1.6-fold). In conclusion, we were able to integrate THP-1-derived iDCs as functional dermal dendritic cell surrogates allowing the qualitative identification of potential sensitizers on the one hand, and drug candidates that potentially suppress sensitization on the other hand in a 3D human skin model corresponding to the 3R principles (“replace”, “reduce” and “refine”).

中文翻译：

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模拟真皮树突细胞激活的人体 3D 免疫能力全层皮肤模型

我们整合了最初由 THP-1 细胞系产生的真皮树突状细胞替代物，作为人类全层皮肤模型中免疫反应的中心介质。因此，THP-1衍生的CD14的敏化剂治疗-, CD11c+未成熟树突状细胞 (iDC) 在 1-氯-2,4-二硝基苯 (DNCB) 存在下导致 p38 MAPK 磷酸化（2.6 倍），并导致抑制剂蛋白 kappa B α (IκBα) 降解NiSO 孵育4（1.6 倍）。此外，NiSO4导致IL-6（2.4倍）、TNF-α（2倍）和IL-8（15倍）的mRNA水平增加。这些结果在蛋白质水平上得到了证实，在 NiSO 存在的情况下对细胞因子释放的影响甚至更强4：IL-8（147 倍）、IL-6（11.8 倍）和 IL-1β（28.8 倍）的细胞因子分泌显着增加。值得注意的是，DNCB 治疗显示 IL-8（28.6 倍）和 IL-1β（5.6 倍）增加。重要的是，NiSO4分离的 iDC 以及作为真皮树突细胞替代物整合到我们的全层皮肤模型 (SM) 中的 iDC 的治疗诱导了分化粘附分子簇 (CD)54 的上调（iDC：1.2 倍；SM：1.3 倍）倍）和共刺激分子和 DC 成熟标记物 CD86（iDC 约 1.4 倍；SM：约 1.5 倍）表面标记物表达。值得注意的是，地塞米松治疗分离的 iDC（CD54：1.3 倍；CD86：2.1 倍）以及组织整合的 iDC（CD54：1.4 倍；CD86：1.6 倍）可抑制 CD54 和 CD86 的表达。折叠）。总之，我们能够将 THP-1 衍生的 iDC 作为功能性真皮树突状细胞替代物进行整合，一方面可以定性鉴定潜在的致敏剂，另一方面可以在相应的 3D 人体皮肤模型中定性鉴定可能抑制致敏的候选药物。 3R 原则（“替换”、“减少”和“优化”）。