División de Inmunología, Facultad de Química, Bioquímica y Farmacia, Universidad Nacional de San Luis, San Luis, Argentina.
Instituto Multidisciplinario de Investigaciones Biológicas (IMIBIO), Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), San Luis, Argentina.
PLoS One. 2018 Mar 1;13(3):e0193573. doi: 10.1371/journal.pone.0193573. eCollection 2018.
Dendritic cells (DCs) play critical functions in the initiation of immune responses. Understanding their role in reactive arthritis (ReA) will help delineate the pathogenesis of this arthropathy. In early studies, we detected IL-12/23p40 deregulation in Yersinia entercolitica (Ye)-induced ReA in TNFRp55-deficient (TNFRp55-/-) mice. In this study, we assessed the contribution of DCs in this overproduction. First, greater levels of IL-12/23p40, IFN-γand IL-17A were confirmed in supernatants of lipopolysaccharide (LPS)-stimulated TNFRp55-/-splenocytes obtained on arthritis onset (day 14 after Ye infection). Later, DCs were identified as a precise source of IL-12/23p40 since increased frequency of splenic IL-12/23p40+DCs was detected in TNFRp55-/- mice. After robust in vivo amplification of DCs by injection of Fms-like tyrosine kinase 3-Ligand (Flt3L)-transfected BL16 melanoma, DCs were purified. These cells recapitulated the higher production of IL-12/23p40 under TNFRp55deficiency. In agreement with these results, TNFRp55-/- DCs promoted Th1 and Th17 programs by co-culture with WT CD4+lymphocytes. A mechanistic study demonstrated that JNK and p38 MAPK pathways are involved in IL-12/23p40 overproduction in purified TNFRp55-/- DCs as well as in the JAWS II cell line. This deregulation was once again attributed to TNFRp55 deficiency since CAY10500, a specific inhibitor of this pathway, compromised TNF-mediated IL-12/23p40 control in LPS-stimulated WT DCs. Simultaneously, this inhibition reduced IL-10 production, suggesting its role mediating IL-12/23p40 regulation by TNFRp55 pathway. These results provide experimental data on the existence of a TNFRp55-mediated anti-inflammatory circuit in DCs. Moreover, these cells may be considered as a novel target in the treatment of ReA.
树突状细胞(DCs)在免疫反应的启动中发挥关键作用。了解它们在反应性关节炎(ReA)中的作用将有助于阐明这种关节病的发病机制。在早期研究中,我们在 TNFRp55 缺陷(TNFRp55-/-)小鼠的耶尔森菌肠结肠炎(Ye)诱导的 ReA 中检测到白细胞介素-12/23p40 的失调。在这项研究中,我们评估了 DCs 在这种过度产生中的贡献。首先,在关节炎发作时(Ye 感染后第 14 天),从脂多糖(LPS)刺激的 TNFRp55-/-脾细胞的上清液中证实了更高水平的白细胞介素-12/23p40、IFN-γ和白细胞介素-17A。后来,由于在 TNFRp55-/-小鼠中检测到脾 IL-12/23p40+DCs 的频率增加,因此确定 DCs 是 IL-12/23p40 的精确来源。通过注射 Fms 样酪氨酸激酶 3 配体(Flt3L)转染的 BL16 黑色素瘤对 DC 进行体内扩增后,分离纯化了 DCs。这些细胞在 TNFRp55 缺陷下重现了更高的白细胞介素-12/23p40 产生。与这些结果一致,TNFRp55-/-DCs 通过与 WT CD4+淋巴细胞共培养促进了 Th1 和 Th17 程序。一项机制研究表明,JNK 和 p38 MAPK 途径参与了纯化的 TNFRp55-/-DCs 中白细胞介素-12/23p40 的过度产生以及 JAWS II 细胞系。由于特定的该途径抑制剂 CAY10500 损害了 LPS 刺激的 WT DCs 中 TNF 介导的白细胞介素-12/23p40 控制,这种失调再次归因于 TNFRp55 缺陷。同时,这种抑制减少了白细胞介素-10 的产生,表明其在 TNFRp55 途径介导白细胞介素-12/23p40 调节中的作用。这些结果提供了关于 DC 中存在 TNFRp55 介导的抗炎回路的实验数据。此外,这些细胞可被视为治疗 ReA 的新靶点。