Samra Z Q, Oguro T, Fontaine R, Ashraf M
Centre for Advanced Molecular Biology, University of the Punjab Lahore, Pakistan.
J Submicrosc Cytol Pathol. 1991 Jul;23(3):379-90.
Monoclonal antibodies (MAb's) to xanthine oxidase (XO) (from bovine milk) were produced by hybridoma technique. Culture supernatants were initially screened using enzyme-linked immunoabsorbant assay (ELISA). Forty four positive clones were subcloned and further characterized by ELISA and immunoblot analysis. Out of fifteen clones, which were positive in immunoblot analysis, one clone N2-26 was also positive in immunocytochemical studies. Indirect immunoperoxidase and enzyme histochemistry staining showed that XO activity is present in endothelial cells of capillaries, small blood vessels and also in interstitial cells. Electron microscopy revealed that diaminobenzidine reaction product was distributed in the cytoplasm of interstitial cells and endothelial cells of capillaries and small blood vessels. This is the first report of the presence of XO in interstitial cells and endothelial cells of small blood vessels. Allopurinol, which inhibits the xanthine oxidase activity, did not have any effect on the immunocytochemical staining. Our results in normal rat heart suggest that XO activity is confined to interstitial cells, endothelial cells of capillaries and small blood vessels.
采用杂交瘤技术制备了针对黄嘌呤氧化酶（XO）（来自牛乳）的单克隆抗体（MAb's）。最初使用酶联免疫吸附测定（ELISA）对培养上清液进行筛选。44个阳性克隆进行了亚克隆，并通过ELISA和免疫印迹分析进一步鉴定。在免疫印迹分析中呈阳性的15个克隆中，一个克隆N2 - 26在免疫细胞化学研究中也呈阳性。间接免疫过氧化物酶和酶组织化学染色显示，XO活性存在于毛细血管、小血管的内皮细胞以及间质细胞中。电子显微镜显示，二氨基联苯胺反应产物分布在间质细胞以及毛细血管和小血管内皮细胞的细胞质中。这是首次报道在小血管的间质细胞和内皮细胞中存在XO。抑制黄嘌呤氧化酶活性的别嘌呤醇对免疫细胞化学染色没有任何影响。我们在正常大鼠心脏中的结果表明，XO活性局限于间质细胞、毛细血管和小血管的内皮细胞。
