Immunochemical detection of the Thomsen-Friedenreich antigen (T-antigen) on the pig lymphocyte plasma membrane

Abstract

A method for the isolation of lymphocytes from pig peripheral blood and preparation of plasma membranes was developed. The method resulted in a ten-fold increase in 5'-nucleotidase activity and neuraminic acid, relative to protein. A cholesterol/phospholipid ratio of 0.93 was obtained. Alkaline borohydride treatment of isolated plasma membrane, after desialylation, released the disaccharide beta-D-galactosyl-(1 leads to 3)-N-acetyl-D-galactosaminitol (the immunodominant group of the T-antigen), which was identified by gas chromatography using a crystalline standard. The disaccharide was not found without prior desialylation, indicating that all the disaccharide units were substituted by neuraminic acid. Serological evidence using the agglutinins from Arachis hypogoea and Helix pomatia confirmed the presence of this disaccharide (T-antigen) in the lymphocyte membrane and indicated that it was the major alkali-labile oligosaccharide of the pig lymphocyte membrane.