Differential localisation and synthesis of porcine factor VIII related antigen (VIIIR:AG) in vascular endothelium and in endothelial cells in culture.
Porcine and human umbilical vein and adult blood vessels were studied for the presence and synthesis of factor VIII related antigen (VIIIR:Ag) and fibronectin (Fn) by immunofluorescence histology and immunoautoradiography. Investigation of human tissue confirmed the widespread distribution of VIIIR:Ag on the endothelium of all blood vessels examined but observations on porcine tissue gave different results. Porcine umbilical vein and porcine adult veins were positively stained for VIIIR:Ag whilst porcine aorta and other pig arteries appeared to be negative or only weakly positive. Some blood vessels (?venous) in the adventitia of porcine aorta were positively stained whilst adjacent ones (?arterial) were negative. Radiolabelled methionine was added to culture medium and proteins synthesised by cultured EC were examined by two dimensional crossed immunoelectrophoresis and autoradiography. Identification of radiolabelled precipitin arcs provided a highly sensitive method for confirming the specificity of antisera and for detecting VIIIR:Ag and Fn. Examination of cultured human umbilical vein EC confirmed the synthesis of VIIIR:Ag and distinguished between VIIIR:Ag and Fn. Studies of porcine umbilical vein EC in culture gave similar results to those observed with corresponding human EC. However, cultures derived from porcine aorta did not demonstrate synthesis of VIIIR:Ag and microscopy failed to locate VIIIR:Ag in these cells with certainty. The results confirmed the synthesis of VIIIR:Ag by human and porcine umbilical vein EC but differences in staining reactions and the apparent inability to synthesise VIIIR:Ag by cells derived from porcine aorta suggested that porcine EC at different anatomical sites may subserve different functions.
D000818:Animals; D000941:Antigens / Q000032:analysis* / Q000096:biosynthesis; D001011:Aorta / Q000032:analysis; D001158:Arteries / Q000032:analysis; D001345:Autoradiography; D002478:Cells, Cultured; D004727:Endothelium / Q000032:analysis / Q000166:cytology; D005169:Factor VIII / Q000032:analysis / Q000096:biosynthesis / Q000276:immunology*; D005353:Fibronectins / Q000032:analysis; D006801:Humans; D007122:Immunoelectrophoresis; D013552:Swine / Q000097:blood*; D014471:Umbilical Veins / Q000032:analysis*; D014841:von Willebrand Factor
D000941:Antigens; D005353:Fibronectins; D014841:von Willebrand Factor; D005169:Factor VIII