Immunohistochemical differentiation between lymphatic vessels and blood vessels--use of anti-basement membrane antibodies and anti-factor VIII-related antigen

Abstract

Several immunohistochemical methods using Factor VIII-Related antigen (FVIIIR:Ag), laminin, Type IV collagen and fibronectin antisera were applied for the purpose of differentiating rat lymphatics from blood vessels by light and electron microscopy. Weibel-Palade bodies (WPB) were demonstrated in both types of vessels by conventional electron microscopy. The immunoreactivity to laminin and Type IV collagen in blood vessels showed a strong, continuous, linear subendothelial staining pattern in contrast to lymphatic vessels in which immunoreactivity was absent or weak in paraffin-embedded sections stained with the indirect immunoperoxidase technique. A positive reaction for fibronectin was observed in all extra-vascular tissue spaces as well as in lymphatics and blood vessels. FVIIIR:Ag and WPB were present in both lymphatic and blood endothelial cells. FVIIIR:Ag antiserum labeled with gold particles was observed only in the vacuoles which were assumed to be identical with WPB as demonstrated by our conventional electron microscopy. We conclude that the immunohistochemical method using laminin and Type IV collagen antisera is a reliable and practical way to differentiate lymphatic vessels from blood vessels by light microscopy.