帕金森病(PD)的特点是神经炎症,多巴胺能神经元的进行性丢失,和α-突触核蛋白(α-Syn)积累成不溶性聚集体,称为路易病理学。61系α-Syn小鼠是已建立的PD临床前模型;Thy-1用于促进人α-Syn表达,散发性PD的特征在9-18个月大时发展。为了加速PD表型,我们将超声处理的人α-Syn预形成的原纤维(PFFs)注入纹状体,在黑质致密质中产生磷酸化Syn(p-α-Syn)包涵体,并显着增加了MHCII类阳性免疫细胞。此外,中脑固有免疫细胞和适应性免疫细胞的浸润和激活增强.然后我们使用了这个新模型,第61行-PFF,为了研究抑制JAK/STAT信号通路的作用,这对于调节先天和适应性免疫反应至关重要。给予JAK1/2抑制剂AZD1480后,免疫荧光染色显示p-α-Syn包涵体和MHCII类表达显着降低。流式细胞术显示CD4+T细胞浸润减少,CD8+T细胞,CD19+B细胞,树突状细胞,巨噬细胞,和内源性小胶质细胞进入中脑。重要的是,单细胞RNA-来自中脑的CD45+细胞的测序分析鉴定出9个小胶质细胞簇,5单核细胞/巨噬细胞(MM)簇,和5个T细胞(T)簇,其中潜在致病性MM4和T3簇与Line61-PFF小鼠的神经炎症反应相关。AZD1480处理减少了细胞数量和抗原提呈基因H2-Eb1,H2-Aa的簇特异性表达,H2-Ab1和Cd74中的MM4簇和促炎基因如Tnf,Il1b,C1qa,和T3集群中的C1qc。一起,这些结果表明,抑制JAK/STAT通路抑制了先天和适应性细胞的活化和浸润,在Line61-PFF小鼠模型中减少神经炎症。
Parkinson\'s disease (PD) is characterized by neuroinflammation, progressive loss of dopaminergic neurons, and accumulation of α-synuclein (α-Syn) into insoluble aggregates called Lewy pathology. The Line 61 α-Syn mouse is an established preclinical model of PD; Thy-1 is used to promote human α-Syn expression, and features of sporadic PD develop at 9-18 months of age. To accelerate the PD phenotypes, we injected sonicated human α-Syn preformed fibrils (PFFs) into the striatum, which produced phospho-Syn (p-α-Syn) inclusions in the substantia nigra pars compacta and significantly increased MHC Class II-positive immune cells. Additionally, there was enhanced infiltration and activation of innate and adaptive immune cells in the midbrain. We then used this new model, Line 61-PFF, to investigate the effect of inhibiting the JAK/STAT signaling pathway, which is critical for regulation of innate and adaptive immune responses. After administration of the JAK1/2 inhibitor AZD1480, immunofluorescence staining showed a significant decrease in p-α-Syn inclusions and MHC Class II expression. Flow cytometry showed reduced infiltration of CD4+ T-cells, CD8+ T-cells, CD19+ B-cells, dendritic cells, macrophages, and endogenous microglia into the midbrain. Importantly, single-cell RNA-Sequencing analysis of CD45+ cells from the midbrain identified 9 microglia clusters, 5 monocyte/macrophage (MM) clusters, and 5 T-cell (T) clusters, in which potentially pathogenic MM4 and T3 clusters were associated with neuroinflammatory responses in Line 61-PFF mice. AZD1480 treatment reduced cell numbers and cluster-specific expression of the antigen-presentation genes H2-Eb1, H2-Aa, H2-Ab1, and Cd74 in the MM4 cluster and proinflammatory genes such as Tnf, Il1b, C1qa, and C1qc in the T3 cluster. Together, these results indicate that inhibiting the JAK/STAT pathway suppresses the activation and infiltration of innate and adaptive cells, reducing neuroinflammation in the Line 61-PFF mouse model.