Ramasesh N, Hastings R C, Krahenbuhl J L
Infect Immun. 1987 May;55(5):1203-6. doi: 10.1128/iai.55.5.1203-1206.1987.
The incorporation of 14C-labeled palmitic acid ( [U-14C]PA) into the phenolic glycolipid-I (PGL-I) fraction of Mycobacterium leprae was studied in a murine macrophage system in vitro. Peritoneal macrophages from Swiss Webster mice were infected with fresh viable or Formalin-killed M. leprae harvested from infected footpads of nu/nu mice, and [U-14C]PA was added to the culture medium. Labeled glycolipid synthesized by live M. leprae was fractionated on a Florisil-silicic acid column and identified as PGL-I by using thin-layer chromatography and localization on a polysulfone membrane with an anti-PGL-I monoclonal antibody. Increased incorporation of [U-14C]PA into the PGL-I fraction was observed in macrophages infected with only live M. leprae. Treatment of the infected macrophages with rifampin caused a significant reduction in the incorporation of palmitic acid into PGL-I. These preliminary studies suggest that PGL-I synthesis can be used to quantitate the metabolism of M. leprae in macrophages in vitro.
在体外小鼠巨噬细胞系统中,研究了14C标记的棕榈酸([U-14C]PA)掺入麻风分枝杆菌酚糖脂-I(PGL-I)组分的情况。用从无胸腺裸鼠感染的足垫中收获的新鲜活的或经福尔马林灭活的麻风分枝杆菌感染瑞士韦伯斯特小鼠的腹腔巨噬细胞,并将[U-14C]PA加入培养基中。由活的麻风分枝杆菌合成的标记糖脂在弗罗里硅土-硅酸柱上进行分离,并通过薄层色谱法以及使用抗PGL-I单克隆抗体在聚砜膜上定位鉴定为PGL-I。在仅感染活的麻风分枝杆菌的巨噬细胞中,观察到[U-14C]PA掺入PGL-I组分增加。用利福平处理感染的巨噬细胞导致棕榈酸掺入PGL-I显著减少。这些初步研究表明,PGL-I合成可用于体外定量巨噬细胞中麻风分枝杆菌的代谢。