De Lemos-Chiarandini C, Alvarez F, Bernard O, Homberg J C, Kreibich G
Hepatology. 1987 May-Jun;7(3):468-75. doi: 10.1002/hep.1840070310.
Human sera, containing anti-liver-kidney microsome antibody as demonstrated by indirect immunofluorescence, were obtained from a subgroup of young patients with autoimmune chronic hepatitis. The anti-liver-kidney microsome antibody-positive sera were used to study the localization of the liver-kidney microsome antigen in hepatocytes. Immunoblot analysis of microsomal subfractions, lysosomal membranes, plasma membranes, mitochondria and purified ribosomes obtained from rat liver demonstrated that this antibody recognizes a protein of 50 kD present only in endoplasmic reticulum membranes. Immunogold labeling of ultrathin frozen sections and immunoperoxidase staining of 11 to 15 micron cryostat sections were used to detect the liver-kidney microsome antigen in rat liver tissue. The anti-liver-kidney microsome antibody binds to antigenic domains on the cytoplasmic face of smooth and rough endoplasmic reticulum membranes of hepatocytes. No labeling was observed of the Golgi apparatus, peroxisomes, mitochondria, lysosomes, nuclei or plasma membranes. Not only was the antigen recognized by the anti-liver-kidney microsome antibody specific for endoplasmic reticulum membranes, but it was also specific for the endoplasmic reticulum of hepatocytes only, since no labeling was observed in any organelle of Kupffer or endothelial cells. Therefore, the anti-liver-kidney microsome antibody can be considered as a marker for endoplasmic reticulum in rat hepatocytes.
通过间接免疫荧光法证实含有抗肝肾微粒体抗体的人血清，取自自身免疫性慢性肝炎的年轻患者亚组。抗肝肾微粒体抗体阳性血清用于研究肝肾微粒体抗原在肝细胞中的定位。对从大鼠肝脏获得的微粒体亚组分、溶酶体膜、质膜、线粒体和纯化核糖体进行免疫印迹分析，结果表明该抗体识别仅存在于内质网膜中的一种50kD的蛋白质。利用超薄冷冻切片的免疫金标记和11至15微米低温恒温器切片的免疫过氧化物酶染色来检测大鼠肝脏组织中的肝肾微粒体抗原。抗肝肾微粒体抗体与肝细胞滑面和粗面内质网膜胞质面的抗原结构域结合。未观察到高尔基体、过氧化物酶体、线粒体、溶酶体、细胞核或质膜有标记。抗肝肾微粒体抗体识别的抗原不仅对内质网膜具有特异性，而且仅对肝细胞的内质网具有特异性，因为在库普弗细胞或内皮细胞的任何细胞器中均未观察到标记。因此，抗肝肾微粒体抗体可被视为大鼠肝细胞内质网的标志物。
