Glycosphingolipids of Membrane Fractions from Normal and Transplanted Canine Kidney

Abstract

Abstract The neutral glycosphingolipids and gangliosides of cell membrane particulate fractions from normal canine kidney and allograft rejected canine kidney have been examined. Those which increased in concentration in the rejected kidney were glucosylceramide, galactosylglucosylceramide, galactosylgalactosylglucosylceramide, N-acetylgalactosaminylgalactosylgalactosylglucosylceramide, N-acetylgalactosaminyl-N-acetylgalactosaminylgalactosylgalactosylglucosylceramide (Forssman hapten), and N-acetylneuraminylgalactosylglucosylceramide. Glycosphingolipids which were lower in the rejected kidney were galactosylceramide, digalactosylceramide, and galactosylsulfatide. The composition of the oligosaccharide moiety of each glycosphingolipid was determined by gas-liquid chromatography. The positions of glycosidic linkages of the sugars were determined by permethylation and gas-liquid chromatographic analysis of the partially methylated alditol acetates. The internal linkages of the oligosaccharides determined were N-acetylgalactosaminyl(1 →3)N-acetylgalactosaminyl(1 →3)galactosyl(1 →4)galactosyl(1 →4)glucosylceramide, galactosyl(1 →4)galactosyl(1 →4)glucosylceramide, and N-acetylneuraminyl(2 →3)-galactosyl(1 →4)glucosylceramide. The glycosphingolipids of canine kidney membrane were found by complement fixation to have little or no antibody binding capacity when tested against serum obtained from transplant recipients after removal of the rejected kidney allograft. Intravenous injection of d-[1-14C]glucosamine into a kidney transplant recipient 4 days after transplantation demonstrated a rapid incorporation into the hexosamine-containing glycosphingolipids of the rejected kidney membrane but not normal kidney membrane. Low levels of radioactivity were incorporated into the gangliosides of both control and rejecting kidneys and into the other neutral glycosphingolipids of control kidneys.