B Lymphoblastoid Cell Lines With Normal and Defective 0 - Glycosylation

Individuals with the T blood group contain terminal serme !threonine-linked N-acetylgalactosamine residues in their blood cells. This is due to lack of UDP-D-galactose: D-N-acetyl galactosamine /9-D-galactosyl transferase from part of their red cells and probably from their leukocytes. We have established B lymphoblastoid cell lines from such an individual by in vitro infection of his lymphocytes with Epstein-Barr virus. The original line contained a mixture of cells reactive and nonreactive with Helix pomatia lectin (Hp). These cells were subcloned after staining with fluorescent Hp by a fluorescence-activated cell sorter (FACS) into homogeneous. phenotypically stable lines of Hppositive (Hp+) and Hp-negative (Hp-) cells. The molecular differences between the membrane glycoproteins were characterized by carbohydrate-specific surface labeling techniques. Hp affinity chromatography. polyacrylamide slab gel electrophoresis and glycopeptide!oligosaccharide