Miliukene V V, Biziuliavichene G Iu, Khaustova L P, Pilinkene A V, Biziuliavichius G A
Tsitologiia. 2007;49(10):853-7.
The fluorometric method was used to study guantitative parameters of phagocytosis of fluorescein-labeled Escherichia coli cells by mouse peritoneal macrophages. E. coli cells were conjugated with fluoresceinisothiocyanate (FITC) and then incubated with macrophages. At the end of the assay phagocytosis was arrested with a lysing solution (0.5% Triton X-100 in 0.01 M phosphate-buffered 0.15 M saline, pH 7.4). Trypan blue at a concentration of 0.04% was used as a quenching agent to differentiate between attachment and ingestion of E. coli cells. The time course analysis within this method showed that phagocytosis of E. coli cells was temperature and opsonin dependent. The number of E. coli cells ingested by macrophages increased rapidly during the initial 60 min of incubation at 37 degrees C. E. coli cells required opsonization with 5% native serum to achieve their optimal uptake. The uptake of nonopsonized bacteria by macrophages was significantly lower that that of opsonized ones (P < 0.05). It was demonstrated that sodium azide inhibited phagocytosis of E. coli cells by mouse peritoneal macrophages in a dose-dependent manner.
采用荧光法研究小鼠腹腔巨噬细胞对荧光素标记的大肠杆菌细胞的吞噬定量参数。将大肠杆菌细胞与异硫氰酸荧光素（FITC）偶联，然后与巨噬细胞一起孵育。在测定结束时，用裂解液（0.01 M磷酸盐缓冲的0.15 M盐水中的0.5% Triton X-100，pH 7.4）终止吞噬作用。浓度为0.04%的台盼蓝用作淬灭剂，以区分大肠杆菌细胞的附着和摄取。该方法的时间进程分析表明，大肠杆菌细胞的吞噬作用依赖于温度和调理素。在37℃孵育的最初60分钟内，巨噬细胞摄取的大肠杆菌细胞数量迅速增加。大肠杆菌细胞需要用5%天然血清进行调理才能实现最佳摄取。巨噬细胞对未调理细菌的摄取明显低于调理细菌（P < 0.05）。结果表明，叠氮化钠以剂量依赖的方式抑制小鼠腹腔巨噬细胞对大肠杆菌细胞的吞噬作用。
