Immunohistochemical localization of xanthine oxidase in human cardiac and skeletal muscle.
The generation of a monoclonal antibody specific to xanthine oxidase and its use in the distribution of the enzyme in human tissue is described. Xanthine oxidase was purified from human and bovine milk by a rapid method, allowing for minimal proteolytic degradation, and the purified enzyme preparations were used for the immunization of BALB/c mice as well as for the subsequent selection of hybridomas. The hybridoma clone X1-7, IgG (2a, kappa-light chain) was selected for further analysis and demonstrated to precipitate xanthine oxidase from human liver and skeletal muscle extracts. As determined by SDS-polyacrylamide gel electrophoresis of eluates from affinity chromatography, the X1-7 antibody bound to a main protein of 155 kDa, from human milk and skeletal muscle, and to proteins of 155, 143 and 95 kDa from human liver. Immunohistochemical studies, using two of the monoclonal antibodies with differing epitope specificity, revealed xanthine oxidase to be localized mainly in the vascular smooth muscle cells but also in a proportion of endothelial cells of capillaries and smaller vessels in both human cardiac and skeletal muscle. Immunoreactivity was additionally observed in human macrophages and mast cells. The results of the present study confirm previous reports of the presence of xanthine oxidase in capillary endothelial cells, but also demonstrates additional localization of the enzyme in vascular smooth muscle cells, macrophages and mast cells. The current findings verify that the distribution of xanthine oxidase in human tissue includes cardiac and skeletal muscle.
Hellsten-Westing
Y
Y
Department of Physiology III, Karolinska Institute, Stockholm, Sweden.
D000818:Animals; D000911:Antibodies, Monoclonal / Q000302:isolation & purification; D006801:Humans; D007150:Immunohistochemistry; D051379:Mice; D008807:Mice, Inbred BALB C; D009131:Muscle, Smooth, Vascular / Q000201:enzymology; D009132:Muscles / Q000201:enzymology*; D009206:Myocardium / Q000201:enzymology; D014969:Xanthine Oxidase / Q000032:analysis*
D000911:Antibodies, Monoclonal; D014969:Xanthine Oxidase