Department of Chemistry, Mass Spectrometry Centre, QOPNA, University of Aveiro, Campus Universitário de Santiago, 3810-193 Aveiro, Portugal.
Centre for Neuroscience and Cell Biology, University of Coimbra 3004-517, Coimbra, Portugal; Faculty of Pharmacy, University of Coimbra 3004-517, Coimbra, Portugal.
Free Radic Biol Med. 2014 Dec;77:217-29. doi: 10.1016/j.freeradbiomed.2014.09.008. Epub 2014 Sep 16.
The pathogenesis of allergic contact dermatitis, the most common manifestation of immunotoxicity in humans, is intimately connected to hapten-induced maturation of dendritic cells (DC). The molecular mechanisms driving this maturational program are not completely known; however, initial danger signals such as the generation of reactive oxygen species (ROS) were shown to play a critical role. Recent evidence linking ROS production, endoplasmic reticulum (ER) stress, and the pathogenesis of several inflammatory diseases led us to analyze, in the present work, the ability of the skin sensitizer 1-fluoro-2,4-dinitrobenzene (DNFB) to evoke ER stress in DC-like THP-1 cells and the concomitant consequences to their immunobiology. We found that DNFB triggers a ROS-dependent activation of the PERK-eIFα-ATF4 unfolded protein response (UPR) branch conferring cytoprotection and modulating the maturation/proinflammatory cell status in a biphasic manner. Early DNFB induction of ATF4 positively modulates autophagy-related genes MAP1LC3B and ATG3 and stabilizes the transcription factor Nrf2, causing a strong induction of the HMOX1-detoxifying gene. Moreover, we observed that in a first phase, DNFB-induced ATF4 upregulates IL8 mRNA levels while blocking CD86, IL1B, IL12B, and CXL10 transcription. Later, following ATF4 decay, HMOX1 and IL8 transcription drastically decrease and CD86, IL1B, and Il12B are upregulated. Overall, our results evidence a connection between sensitizer-induced redox imbalance and the establishment of ER stress in DC-like cells and provide new insights into the role of UPR effectors such as ATF4 to the complex DC maturational program.
变应性接触性皮炎的发病机制与树突状细胞(DC)的半抗原诱导成熟密切相关,是人类免疫毒性最常见的表现。尽管推动这一成熟程序的分子机制尚不完全清楚,但最初的危险信号,如活性氧(ROS)的产生,已被证明起着关键作用。最近有证据将 ROS 产生、内质网(ER)应激与几种炎症性疾病的发病机制联系起来,促使我们在本工作中分析皮肤敏化剂 1-氟-2,4-二硝基苯(DNFB)在 DC 样 THP-1 细胞中引发 ER 应激的能力及其对其免疫生物学的伴随后果。我们发现,DNFB 触发依赖 ROS 的 PERK-eIFα-ATF4 未折叠蛋白反应(UPR)分支的激活,赋予细胞保护作用,并以双相方式调节成熟/促炎细胞状态。早期 DNFB 诱导的 ATF4 正向调节自噬相关基因 MAP1LC3B 和 ATG3,并稳定转录因子 Nrf2,导致 HMOX1 解毒基因的强烈诱导。此外,我们观察到在第一阶段,DNFB 诱导的 ATF4 上调 IL8 mRNA 水平,同时阻断 CD86、IL1B、IL12B 和 CXL10 的转录。随后,在 ATF4 衰减后,HMOX1 和 IL8 的转录急剧下降,而 CD86、IL1B 和 Il12B 上调。总之,我们的研究结果表明,敏化剂诱导的氧化还原失衡与 DC 样细胞中 ER 应激的建立之间存在联系,并为 UPR 效应物(如 ATF4)在复杂的 DC 成熟程序中的作用提供了新的见解。