Department of Pharmacology, Medical University, Bialystok, Poland.
Adv Exp Med Biol. 2019;1133:55-63. doi: 10.1007/5584_2018_323.
Exposure to ambient particulate matter (PM) increases mortality and morbidity due to respiratory and cardiovascular diseases. The aim of this study was to assess the effect of standardized urban dust (UD) on phagocytosis and autophagy in a monocyte-macrophage cell line (THP-1 cells). The cells were grown for 24 h in the medium supplemented with 200 μg·mL coarse carbon black (CB) or UD. In some experiments glutathione (GSH) was depleted in THP-1 cells by buthionine sulfoximine. The cells were double stained with green latex beads (phagocytosis) and with red autophagy marker (LC3) and were evaluated in a flow cytometer. In naïve THP-1 cells, about 61% of them were classified as "negative", while 39% were classified as "double-positive". Both GSH depletion and UD treatment produced three distinct subpopulations of cells on bivariate scatterplots. A new subpopulation of cells (about 24% of the total number) appeared, with a lower autophagy and phagocytosis, but with a higher autophagy/phagocytosis ratio, when compared to highly positive cells. CB affected, to some extent, phagocytosis without a substantial effect on autophagy. In conclusion, the research on distinct pathways of immune cell activation may be relevant to the diagnostics and therapy of PM-induced pneumotoxicity, inflammation, and tumorigenesis.
暴露于环境颗粒物（PM）会增加因呼吸道和心血管疾病导致的死亡率和发病率。本研究旨在评估标准化城市尘埃（UD）对单核细胞-巨噬细胞系（THP-1 细胞）吞噬作用和自噬作用的影响。将细胞在补充有 200μg·mL 粗碳黑（CB）或 UD 的培养基中生长 24 小时。在一些实验中，通过丁硫氨酸亚砜胺使 THP-1 细胞中的谷胱甘肽（GSH）耗竭。用绿色乳胶珠（吞噬作用）和红色自噬标志物（LC3）对细胞进行双重染色，并在流式细胞仪中进行评估。在未成熟的 THP-1 细胞中，约 61%的细胞被归类为“阴性”，而 39%的细胞被归类为“双阳性”。GSH 耗竭和 UD 处理均在双变量散点图上产生了三个不同的细胞亚群。当与高阳性细胞相比，出现了一个新的细胞亚群（约占总数的 24%），其自噬和吞噬作用均较低，但自噬/吞噬作用比值较高。CB 在一定程度上影响吞噬作用，但对自噬作用没有实质性影响。总之，对免疫细胞激活的不同途径的研究可能与 PM 诱导的肺毒性、炎症和肿瘤发生的诊断和治疗有关。
