背景:帕金森病(PD)是一种慢性神经退行性疾病,其特征是黑质中多巴胺能神经元的丢失,促进中枢神经系统持续的炎症环境。调节性T细胞(Tregs)在控制炎症中起重要作用,并可能发挥神经保护作用。的确,在PD中已经报道了Treg数量和功能的减少。在这种情况下,普拉克索,一种用于治疗PD症状的多巴胺能受体激动剂,已被证明可以增加Treg细胞的外周水平并改善其抑制功能。这项工作的目的是确定普拉克索对免疫调节Treg细胞的作用及其对从人胚胎干细胞分化的人多巴胺能神经元的神经保护作用。
方法:从健康人供体的白细胞中分选Treg细胞。用2或200ng/mL浓度的普拉克索处理的CD3/CD28活化和未活化的Treg细胞进行测定。在细胞毒性测定中,用6-羟基多巴胺(6-OHDA)将这些调节细胞与体外分化的人多巴胺能神经元共培养。通过将激活的产生IL-10的Treg细胞与神经元共培养来研究白介素-10(IL-10)的作用。为了进一步研究治疗对Tregs的影响,普拉克索治疗的基因表达,通过Fluidigm分析测定CD3/CD28活化的Treg细胞。
结果:普拉克索处理的CD3/CD28激活的Treg细胞在用6-OHDA攻击时对多巴胺能神经元显示出显著的保护作用。普拉克索处理的活化的Treg细胞通过涉及IL-10释放和与调节和神经保护相关的基因激活的机制显示出神经保护能力。
结论:抗CD3/CD28激活的Treg细胞保护多巴胺能神经元免受6-OHDA诱导的损伤。此外,激活,产生IL-10,普拉克索治疗的Tregs也诱导了神经保护作用,和这些共培养物的上清液促进轴突生长。普拉克索治疗,激活的Tregs以浓度依赖的方式改变了它们的基因表达,并增强了TGFβ相关的多巴胺受体调节和免疫相关途径。这些发现为开发用于治疗PD的免疫调节疗法开辟了新的视角。
BACKGROUND: Parkinson\'s disease (PD) is a chronic neurodegenerative disorder characterized by the loss of dopaminergic neurons in the substantia nigra, which promotes a sustained inflammatory environment in the central nervous system. Regulatory T cells (Tregs) play an important role in the control of inflammation and might play a neuroprotective role. Indeed, a decrease in Treg number and function has been reported in PD. In this context,
pramipexole, a dopaminergic receptor agonist used to treat PD symptoms, has been shown to increase peripheral levels of Treg cells and improve their suppressive function. The aim of this work was to determine the effect of
pramipexole on immunoregulatory Treg cells and its possible neuroprotective effect on human dopaminergic neurons differentiated from human embryonic stem cells.
METHODS: Treg cells were sorted from white blood cells of healthy human donors. Assays were performed with CD3/CD28-activated and non-activated Treg cells treated with
pramipexole at concentrations of 2 or 200 ng/mL. These regulatory cells were co-cultured with in vitro-differentiated human dopaminergic neurons in a cytotoxicity assay with 6-hydroxydopamine (6-OHDA). The role of interleukin-10 (IL-10) was investigated by co-culturing activated IL-10-producing Treg cells with neurons. To further investigate the effect of treatment on Tregs, gene expression in
pramipexole-treated, CD3/CD28-activated Treg cells was determined by Fluidigm analysis.
RESULTS: Pramipexole-treated CD3/CD28-activated Treg cells showed significant protective effects on dopaminergic neurons when challenged with 6-OHDA.
Pramipexole-treated activated Treg cells showed neuroprotective capacity through mechanisms involving IL-10 release and the activation of genes associated with regulation and neuroprotection.
CONCLUSIONS: Anti-CD3/CD28-activated Treg cells protect dopaminergic neurons against 6-OHDA-induced damage. In addition, activated, IL-10-producing, pramipexole-treated Tregs also induced a neuroprotective effect, and the supernatants of these co-cultures promoted axonal growth. Pramipexole-treated, activated Tregs altered their gene expression in a concentration-dependent manner, and enhanced TGFβ-related dopamine receptor regulation and immune-related pathways. These findings open new perspectives for the development of immunomodulatory therapies for the treatment of PD.