Kiss A L, Röhlich P
Eur J Cell Biol. 1984 May;34(1):88-95.
The uptake mechanism of homologous IgG and immune complex, and the participation of coated vesicles in this process were studied in rat peritoneal macrophages. Peroxidase-antiperoxidase (PAP) immune complex produced in rat, and purified rat IgG adsorbed to gold particles (IgG-Au) were used as ligands. Freshly collected peritoneal macrophages were preincubated with the ligands at 4 degrees C, washed, warmed up to 37 degrees C, maintained in a serum-free culture medium for 5 sec to 30 min and subsequently fixed for electron microscopy. In the IgG-Au experiments, acid phosphatase reaction was also applied to identify lysosomes, and ruthenium red to trace membranes exposed to the extracellular space. At the end of the preincubation period PAP and IgG were found randomly distributed on the external surface of the plasma membrane. After warming up the cells to 37 degrees C, the ligands bound to the plasma membrane showed a tendency to move towards deep labyrinthic invaginations of the cell surface from where they were internalized via coated pits and coated vesicles. In the initial period, these structures seemed to be the primary carriers of the ligands. In the period between 5 and 10 min, ligands were concentrated in vacuoles (endosomes) located in the deeper cytoplasm, while after 30 min, they were present in large lysosome-like or multivesicular bodies, which were found to be acid phosphatase positive.
在大鼠腹膜巨噬细胞中研究了同源IgG和免疫复合物的摄取机制,以及有被小泡在此过程中的参与情况。用大鼠产生的过氧化物酶-抗过氧化物酶(PAP)免疫复合物以及吸附到金颗粒上的纯化大鼠IgG(IgG-Au)作为配体。将新鲜收集的腹膜巨噬细胞在4℃下与配体预孵育,洗涤后,升温至37℃,在无血清培养基中维持5秒至30分钟,随后固定用于电子显微镜观察。在IgG-Au实验中,还应用酸性磷酸酶反应来鉴定溶酶体,并用钌红来追踪暴露于细胞外空间的膜。在预孵育期结束时,发现PAP和IgG随机分布在质膜的外表面。将细胞升温至37℃后,与质膜结合的配体显示出从细胞表面向深层迷路状内陷移动的趋势,它们通过有被小窝和有被小泡从那里内化。在最初阶段,这些结构似乎是配体的主要载体。在5至10分钟之间,配体集中在位于较深细胞质中的液泡(内体)中,而在30分钟后,它们存在于大的溶酶体样或多泡体中,发现这些结构呈酸性磷酸酶阳性。