Jeanneau C, Sultan Y
J Histochem Cytochem. 1982 Nov;30(11):1091-6. doi: 10.1177/30.11.6815260.
Factor VIII/von Willebrand factor antigen (VIII R:Ag) has been localized in human endothelial cells by an immunoperoxidase method and electron microscopy. Specific antibodies against VIII R:Ag were isolated from monospecific rabbit antisera by an immunoadsorbant. Fab fragments were prepared and coupled to peroxidase. Native human endothelial cells from umbilical cord veins were incubated with the Fab conjugates. Peroxidase labeling was seen on the plasma membrane, in vesicles, and in the Golgi apparatus. In order to eliminate nonspecific antibody internalization, the same procedure was reproduced on fixed endothelial cells and similar localization was observed. Staining was only found when specific VIII R:Ag Fab conjugates were used. All the controls were devoid of labeling, thus the peroxidase reaction product localized in the Golgi cisternae is specific for VIII R:Ag and suggests VIII R:Ag synthesis in endothelial cells.
采用免疫过氧化物酶法和电子显微镜技术，已将凝血因子VIII/血管性血友病因子抗原（VIII R:Ag）定位在人内皮细胞中。通过免疫吸附剂从单特异性兔抗血清中分离出针对VIII R:Ag的特异性抗体。制备Fab片段并与过氧化物酶偶联。将来自脐带静脉的天然人内皮细胞与Fab缀合物一起孵育。在质膜、小泡和高尔基体中可见过氧化物酶标记。为了消除非特异性抗体内化，在固定的内皮细胞上重复相同的程序，并观察到类似的定位。仅当使用特异性VIII R:Ag Fab缀合物时才发现染色。所有对照均无标记，因此高尔基体池中定位的过氧化物酶反应产物对VIII R:Ag具有特异性，提示内皮细胞中VIII R:Ag的合成。
