Kays Matthew, Kelly Anna, McGinnis Farrah, Woods Clara, Brown Candice, Coulibaly Aminata P
Department of Neuroscience, Rockefeller Neuroscience Institute, West Virginia University.
Morrisey School of Arts and Science, Boston College.
bioRxiv. 2025 Jul 24:2025.04.21.649766. doi: 10.1101/2025.04.21.649766.
Recent reports show that neutrophil activity plays a role in the cognitive decline associated with Alzheimer's Disease (AD). There is evidence of altered functions in neutrophils isolated from AD patients. Whether these altered functions are inherent to the AD disease state is unknown. The goal of this study was to determine if neutrophil functions are altered in AD mice and if these changes occur only after symptoms appear. To address this hypothesis, we used a primary neuronal culture model, generated from 3xTg perinatal mice, since AD is considered a CNS disease. The 3xTg primary neuronal culture gradually increase the release of Aβ (40 and 42) as the culture ages. To assess neutrophil functions, neutrophils isolated from young male/female mice (3-6 months of age) or aged male/female mice (16-18 months of age) of WT or 3xTg mice were exposed to 3xTg primary neuronal cultures. To assess phagocytosis, we characterized the effect of neutrophils on pathogenic amyloid beta (Aβ) 42 levels. To assess the levels of immune modulators (cytokines, chemokines, growth factors, NETosis, and neutrophil granular content), culture media were assessed using Luminex multiplex assay. Our results show that neutrophils from young AD mice have impaired phagocytosis, as observed in a decreased ability to remove Aβ and cellular debris in vitro. Neutrophils from young AD mice also increased release of pro-inflammatory cytokines, granule content, and NETs in 3xTg primary neuronal cultures. Interestingly, neutrophils from aged 3xTg mice decreased Aβ levels in culture and expression of proinflammatory cytokines when compared to neutrophils from aged WT mice. These neutrophils increased their release of granule content and NETs in 3xTg primary neuronal cultures. These data show that in AD neutrophil function is altered both prodromal (young mice) and diseased (old mice) stage.
最近的报告显示，中性粒细胞活性在与阿尔茨海默病（AD）相关的认知衰退中起作用。有证据表明，从AD患者分离出的中性粒细胞功能发生了改变。这些功能改变是否是AD疾病状态所固有的尚不清楚。本研究的目的是确定AD小鼠的中性粒细胞功能是否发生改变，以及这些变化是否仅在症状出现后才发生。为了验证这一假设，我们使用了由3xTg围产期小鼠产生的原代神经元培养模型，因为AD被认为是一种中枢神经系统疾病。随着培养时间的延长，3xTg原代神经元培养物中Aβ（40和42）的释放逐渐增加。为了评估中性粒细胞功能，将从野生型（WT）或3xTg小鼠的年轻雄性/雌性小鼠（3 - 6个月龄）或老年雄性/雌性小鼠（16 - 18个月龄）分离出的中性粒细胞暴露于3xTg原代神经元培养物中。为了评估吞噬作用，我们研究了中性粒细胞对致病性淀粉样β（Aβ）42水平的影响。为了评估免疫调节剂（细胞因子、趋化因子、生长因子、中性粒细胞胞外陷阱形成和中性粒细胞颗粒内容物）的水平，使用Luminex多重分析法评估培养基。我们的结果表明，年轻AD小鼠的中性粒细胞吞噬功能受损，体外清除Aβ和细胞碎片的能力下降。年轻AD小鼠的中性粒细胞在3xTg原代神经元培养物中还增加了促炎细胞因子的释放、颗粒内容物和中性粒细胞胞外陷阱的形成。有趣的是，与老年WT小鼠的中性粒细胞相比，老年3xTg小鼠的中性粒细胞降低了培养物中的Aβ水平和促炎细胞因子的表达。这些中性粒细胞在3xTg原代神经元培养物中增加了颗粒内容物和中性粒细胞胞外陷阱的释放。这些数据表明，在AD中，中性粒细胞功能在疾病前驱期（年轻小鼠）和疾病期（老年小鼠）均发生改变。
