Renier G, Rousselet M C, Carrere F, Croue A, André C, Oksman F, Chevailler A, Hurez D
Laboratoire d'Immunopathologie, CHU, Créteil, France.
J Autoimmun. 1994 Feb;7(1):133-43. doi: 10.1006/jaut.1994.1010.
Autoantibodies directed against cellular constituents rarely react with antigens localized in the Golgi apparatus and little information is available regarding these particular antibodies. Although thousands of samples have been examined for autoantibodies in our laboratory on a routine basis, only three human sera with anti-Golgi antibodies could be studied. Using pre-embedding immunoelectron microscopy we have demonstrated that these sera have antibodies reacting with antigens located in the Golgi apparatus. The reaction product was exclusively located on cisternal and vesicular Golgi membranes. No intraluminal staining was seen and some saccules were negative. No specificity for a peculiar tissue or cell line was noted, suggesting that the targets or these autoantibodies are evolutionarily conserved. The F(ab')2 fragments retained full binding capacity in indirect immunofluorescence experiments, confirming true antibody activity. When tested by immunoblotting, the three sera reacted with different antigens with relative molecular weights of respectively 230, 150 and 80 kDa. The antigens recognized by anti-Golgi antibodies in two of the three sera were insensitive to trypsin degradation. Together, these results suggest that a set of different autoantigens are recognized by sera from various patients.
针对细胞成分的自身抗体很少与定位于高尔基体的抗原发生反应,关于这些特定抗体的信息也很少。尽管我们实验室已常规检测了数千份样本中的自身抗体,但仅有三份含抗高尔基体抗体的人血清可供研究。利用包埋前免疫电子显微镜技术,我们已证实这些血清中的抗体可与位于高尔基体的抗原发生反应。反应产物仅位于高尔基体的扁平囊和囊泡膜上。未见腔内染色,部分扁平囊呈阴性。未观察到对特定组织或细胞系的特异性,这表明这些自身抗体的靶标在进化上是保守的。在间接免疫荧光实验中,F(ab')2片段保留了完全的结合能力,证实了其真正的抗体活性。通过免疫印迹法检测时,这三份血清分别与相对分子质量为230、150和80 kDa的不同抗原发生反应。在三份血清中的两份中,抗高尔基体抗体识别的抗原对胰蛋白酶降解不敏感。这些结果共同表明,不同患者的血清识别一组不同的自身抗原。